How are Tissue Samples Processed for Histology?

There are numerous steps involved in preparing tissue samples for histology. Fixed tissue cannot simply be taken and embedded. This article focuses on the in-between stage of processing tissue for histology. These six steps occur between tissue fixation and the embedding or sectioning processes.

Six Steps in Tissue Processing for Histology

Step 1: Labeling Cassettes

After fixation, tissue samples are transferred to a tissue cassette. These come in a variety of sizes. They hold and protect tissue during the process. Once the embedding stage has been reached, the cassette lid is removed. The primary part of the cassette forms a base for the paraffin wax block. Cassettes have to be properly labeled either by hand or using a cassette labeling machine. The three steps in processing tissue include dehydrating, clearing, and infiltrating. Each of these steps involves diffusing the solution into tissue and dispersing the previous solution.

Step 2: Carousel Machine

Dedicated tissue processing machines carry out tissue processing for histology in most of today’s modern labs and institutes.  Older machines were designed similarly to a carousel containing a cage where the cassettes were placed. Inside the carousel, glass beakers contain solvents and solutions that ensure the tissue is dehydrated, cleared, and ready for embedding in paraffin wax. The carousel agitates the cage in each solution and continues moving through the various solutions in the dehydration and clearing method. Modern processors contain a chamber that holds specimens while the various solutions are pumped in and out of the chamber. Generally, the process takes about six hours and most labs set it up to run overnight.

Step 3: Remove the Water

The tissue samples need to be dehydrated first. The water may be bound to the tissue or free. Paraffin wax is hydrophobic, so the water has to be removed before the tissue can be infiltrated with wax. The water is carried out by immersing the tissue into a series of ethanol solutions. The series of solutions contain increasing concentrations of ethanol until a 100% water-free alcohol is achieved. The increasing concentrations ensure the water in the tissue is slowly replaced by alcohol and to avoid distortion of the tissue. During the process, some components of the cell are removed.

Step 4: Ethanol Won’t Mix with Wax

Even though the tissue reaches the final stage of dehydration in ethanol, it cannot proceed directly to wax embedding. Ethanol and wax will not mix. Tissue clearing is necessary. Clearing refers to the solvents used. They have a high refractive index. When tissue is immersed in the solvents, it becomes clear and transparent.

Step 5: Clearing Agents

Xylene is a solvent that is used in the intermediate stage. This clearing agent is used since it is miscible with both paraffin wax and ethanol. After dehydration, the tissue is immersed in xylene solutions until it gradually replaces the ethanol. Once the tissue is embedded, the xylene is replaced by molten paraffin wax. During these stages, the tissue may shrink because xylene also removes fat residue left in the tissue samples.

Step 6: The Tissue Block

The final stage is called blocking out or infiltration. During infiltration, the last xylene is replaced with molten wax which infiltrates the tissue. Three different wax immersions are used to make sure no clearing agent is left in the tissue sample. After the last infiltration, tissue cassettes removed from the cassettes and placed into a metal mold which is filled with molten wax. The mold is transferred to the cold plate and allowed to set. Once the tissue is set it is ready for the journey of sectioning, making slides, and immunohistochemistry.

In Conclusion

There are many steps necessary to properly prepare tissue samples for histology. The steps must be carried out in a specific order with no deviations to ensure consistency among tissue samples.

References

https://webpath.med.utah.edu/HISTHTML/HISTOTCH/HISTOTCH.html

https://www.leicabiosystems.com/knowledge-pathway/an-introduction-to-specimen-processing/

https://bitesizebio.com/13398/how-histology-slides-are-prepared/

https://www.researchgate.net/publication/233844919_Routine_Histotechniques_Staining_and_Notes_on_Immunohistochemistry

Preparation, Storage, and Applications of FFPE Tissue

FFPE (Formalin-Fixed Paraffin-Embedded) tissue specimens have played an integral role in research and therapeutic applications for decades. FFPE is the way biopsy specimens are preserved and prepared. The process ensures the specimens are suitable for examination, diagnostic/drug development, and experimental research. Once a tissue sample has been obtained, it is preserved. This process includes fixing it in formaldehyde, or formalin. It ensures the proteins and vital structures in the sample are preserved. After the tissue is preserved intact, it is embedded in a paraffin block. This process makes it easier to slice off the right size, so it fits on a microscopic slide for examining.

Methods of Tissue Collection

There are a variety of methods used for collecting tissue specimens. They can be obtained from both normal and diseased individual donors. Another method is to match a pair of samples from one person. This involves taking a sample of healthy tissue and one of diseased tissue from the same individual. Oncologists often use these types of samples for comparison. They can observe a primary tumor with a more distant metastatic tumor in one patient.

Tissue Sample Sources

Tissue is often excised for patients or donors. But it can also be obtained from other animals including mice or snakes. The block only has to measure a few centimeters, but the actual size may be determined by the tissue source and the nature of extraction. Immediately after excision, the tissue is immersed in a solution of 10% neutral-buggered formalin where it remains for 18 to 24 hours. This helps to harden the tissue so can endure the next steps.

FFPE Tissue Sample Preparation

The tissue must be dehydrated and cleared before being infiltrated with wax. This is often done using concentrates of ethanol. After it is dehydrated, it is embedded into IHC-grade wax or paraffin. Timing of fixation is vitally important to FFPE. If it is fixed too soon it can be useless for molecular biology studies. But they do need to be fixed long enough to preserve them. Mishandling samples can result in misleading deductions.

The method of FFPE tissue preparation depends on requirements as they are outlined by physicians or researchers who are requesting the tissues. They may have specifications about the size, purpose, or how it is cut. One example is muscle tissue which may be cut along the grain of the muscle fibers, or across the fibers. Certified medical pathologists are often involved in preparing samples to ensure the procedure is accurate, and to assess quality.

Storage of Prepared Tissue Samples

Once tissue samples have been completed, they are stored in a tissue bank, research center, or biorepository. Records are kept meticulously as to when tissue was collected and preserved. Other information such as the age, ethnicity, and gender of the donor as well as the origin, duration and the stage of diseased tissue are recorded. There are guidelines that must be followed such as obtaining signed consent forms and other legal documents. These can impact whether FFPE tissue can be used in clinical trials or in research. Once FFPE tissue slides are prepared properly, they are durable and can be stored at room temperature for years.

Applications of FFPE Tissue

FFPE tissue is often used in immunohistochemistry or IHC. This technique involves mounting the tissue sections on a slide, then they are covered with a solution that contains antibodies. The antibodies bind to specific structures or proteins. Then, stains are used to make the antibodies visible. This shows the types of structures or proteins are in the tissue sample and where they are located. Information like this is critically important in the medical field when doctors are looking for signs of diseases. IHC information is vital to many cancer research. Other applications include:

·   Oncology – FFPE tissues are essential to the study of oncology or cancer.

·   Hematology – FFPE tissue is vital in the study of blood and related disorders. Hematology has been key to discovering many cures for diseases caused by anomalies in the blood and its components.

·   Immunology – Tissue samples from individuals who have an autoimmune disease help determine a cause and develop medication for treatment.

·   Comparative – Tissue samples from healthy donors is beneficial for comparing with diseased tissue samples. This is imperative for research and development.

In Conclusion

It is certain that there is future work to be done using FFPE tissue samples. They provide a source of RNA, DNA, and proteins beneficial to medical research. The process must be protected to ensure high-quality standards are observed to ensure research and results are untainted.

Resources:

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4077003/

https://pubmed.ncbi.nlm.nih.gov/18711211/

https://www.nature.com/articles/448959a

https://audubonbio.com/tissues/ffpe-tissue/

https://meridian.allenpress.com/aplm/article/138/11/1520/128727/A-Review-of-Preanalytical-Factors-Affecting

https://austinpublishinggroup.com/proteomics/fulltext/ap-v1-id1002.php

https://www.future-science.com/doi/10.2144/000114414

https://www.tandfonline.com/doi/full/10.1080/10520295.2018.1446101

How Cryopreservation is used to Preserve Cells and Tissue

Cryopreservation is the method used to preserve cells and tissue by freezing them. The specialized process prevents intracellular ice crystals and dehydration.  Either of these can destroy organelles or cause the cell to die while undergoing the freezing process.

Cryopreservation Agents

Dimethyl sulfoxide (DMSO) and glycerol are two agents used in the cryopreservation process. Glycerol is used for freezing red blood cells. DMSO is used to help protect most other types of tissues and cells. Organisms that can survive extreme dehydration have a sugar called trehalose. This sugar is used in freeze-drying. Trehalose can stabilize cell membranes. It is also useful for helping to preserve sperm, blood cells, and stem cells.

Cellular Cryopreservation Systems

In most cases, cryopreservation is accomplished using a controlled-rate freezer.  This system delivers liquid nitrogen into a closed chamber that houses the cell. The process requires close monitoring to prevent ice crystal formation and dehydration. The cells typically go from room temperature to -130 degrees Fahrenheit (-90 Celsius). Once the cells are frozen, they are transferred to a liquid nitrogen freezer that maintains these types of extremely cold temperatures.

Cryopreservation Applications

One of the most important applications of cryopreservation is freezing and storing hematopoietic stem cells. These cells are in peripheral blood and bone marrow. Before a patient undergoes high-dose chemotherapy, hematopoietic stem cells are retrieved from their bone marrow. After they complete their treatment, their cells which were cryopreserved are thawed so they can be infused back into their body. High-dose chemotherapy is toxic to bone marrow. Being able to cryopreserve hematopoietic stem cells enhances the treatment outcome for some types of solid tumor malignancies and lymphomas.

Patients with leukemia have cancerous blood cells and can’t be used for autologous bone-marrow rescue.  Therefore, leukemia patients rely on cryopreserved blood collected from other sources such as stem cell donors or cryopreserved blood collected from the umbilical cords of newly born infants.

Research continues, but it’s been widely accepted since the 1990s that hematopoietic stem cells and mesenchymal stem cells can be differentiated into nerve tissue, bone, cardiac muscle tissues, and skeletal tissue. Tissue culture systems remain an area of intense study and research as well as cryopreservation of these types of cells.  There is hope that in the future these cells can be used for many applications including disorders of muscle systems, nervous disorders, and diseases of the heart and liver.

Other Uses of Cryopreservation

Cryopreservation is used to freeze and store sperm and human embryos as well. IVF (in vitro fertilization) relies on the freezing of extra embryos. Couples can choose to use cryopreserved embryos if IVF fails with fresh embryos. Frozen embryos are thawed and then implanted into the uterus.

Storage of Frozen Tissue and Cells

When cells are properly frozen, they can live for more than a decade. Some tissues can also be cryopreserved successfully such as veins, aortic tissue, cardiac valves, and parathyroid glands. Freezing has long been used to store and maintain long-term viability of sperm, ova, and early human embryos. Freezing procedures for preserving these types of tissue are well-established.  

Conclusion

Cryopreservation has stood the test of time and plays an important role in the treatment of some cancers and diseases. As research continues, more opportunities to use frozen tissues and cells will become apparent. The process of cryopreservation is certain to continue being an integral part of medical research and treatment on many levels.

References

https://www.biocompare.com/Bench-Tips/560400-Top-Tips-for-Freezing-and-Thawing-Cells-to-Maintain-Viability/

https://pubmed.ncbi.nlm.nih.gov/18080461/

https://www.sciencedirect.com/science/article/abs/pii/S0304416520302609

The Importance of On-Site Clinical Trial Monitoring

There are three main types of clinical monitoring including on-site, remote, and centralized. Remote and centralized monitoring are often confusing since they are both conducted away from the site of the clinical trial. On-site monitoring, on the other hand, involves an in-person evaluation carried out at the site.

Purpose of On-Site Clinical Trial Monitoring

With the latest digitization and technological advancements, more of the monitoring processes can be facilitated remotely. However, when risk-based quality management involves regulatory focus on-site monitoring provides an undeniable value to a clinical study. Monitoring provides three purposes:

·   Protect the rights and well-being of human subjects

·   Compliance with protocols and applicable regulatory requirements

·   Verify the accuracy and completeness of the data

On-site clinical trial monitoring helps achieve these three goals and provides validation which is valuable to future clinical research.

Advantages of On-Site Clinical Trial Monitoring

Build Relationships with the On-Site Study Team

Even though it may seem like a small thing, there are huge benefits to the two stakeholders (study team and monitors) who build a relationship. The monitor essentially works as a mediator between companies and the study team. The monitor often mediates questions and answers between the sponsor and the site. Therefore, the monitor offers a lot of support to the trial. When a successful relationship is forged, the monitor is able to detect and resolve issues at early stages. Ideally, the monitor and site will experience close collaboration.

Checking for Data and Knowledge Accuracy

Technology presents a wide range of options when it comes to remote monitoring. Some elements such as accuracy, compliance, and quality can be checked remotely. But the presence of a monitor on the site is necessary for some trials. Some examples where remote monitoring is not adequate include:

·   Source Data Verification (SDV) – Due to measures used to protect patient information, some files may not be accessed remotely.

·   Handling Investigational Medicinal Product (IMP) – In most instances, proper handling of the IMP (storage, distribution, shipping, destruction) needs strict surveillance on-site.

·   Patient Information & Informed Consent – To ensure patient safety and accordance with regulations, the entire process of informed consent should be followed.

·   Patient Enrollment – A monitor helps ensure that subjects included in the trial meet the inclusion or exclusion criteria. An on-site monitor can check paper and electronic records to ensure compliance with the eligibility criteria.

·   Adverse Event Reporting – Monitors can assure that no adverse events are missing or unreported.

·   Investigator Site File (ISF) – Site monitors ensure continuous traceability over the course of the trial and check all essential documents for completeness and solidity.

·   Case Familiarity – The study team must have great working knowledge and understanding of the trial in order to ensure compliance with the protocol, patient safety, and data integrity. Direct contact between the monitor and study team ensures proper study conduct through implemented processes and procedures.

Immediate Reaction and Solutions

On-site monitors can mean immediate actions and quick solutions to any problems or issues that occur. This encourages staff to effectively optimize their processes and closely collaborate with on-site monitors to ensure a smooth study.

Final Thoughts

Perhaps there is not one single monitoring approach that works best for every type of clinical study or study setting. Monitoring strategies may need to be adapted and planned by taking account of the characteristics and potential risks specific to each study. Of the different types of clinical trial monitoring, on-site monitors offer hands-on, real-time monitoring that is effective and efficient.

References:

https://www.lexitas.com/wp-content/uploads/sites/6/2017/10/The-Hidden-Value-of-Onsite-Monitoring.pdf

https://journals.sagepub.com/doi/full/10.1177/1747016120933923#:~:text=Monitoring%20of%20clinical%20trials%20is,principles%20of%20good%20clinical%20practice.

http://www.jirb.org.tw/DB/File/Download/970127-05_Monitor%20ResponsibilitiesTaipei_Christine%20Maure.pdf

https://globalhealthtrials.tghn.org/articles/clinical-trial-monitoring/

https://www.fda.gov/media/116754/download

https://cheos.ubc.ca/research-in-action/clinical-trial-monitoring-101/

https://khpcto.co.uk/SOPs/03_MonitoringSOP.php

Has COVID-19 Affected the Handling of Biospecimens?

The collection of biospecimens continue even during the midst of the Coronavirus pandemic. Even though there is a global concern about COVID-19, laboratories, experimentation, and sample collection must continue. Some biorepositories and laboratories will turn to focus on the collection and examination of Covid-related biospecimens, others will continue to forge ahead trying to make a difference in their fields of influence. Research must continue as patients are not exempt from serious diseases and conditions due to the pandemic. Repositories must continue collecting tissue and cell samples and making them available for research purposes. But how does the Covid pandemic affect biospecimen collecting, storage, and distribution?

Handling Biospecimens

In general, guidelines for the safe handling of biospecimens are in place and do not require further recommendations. Precautions are in place for the purpose of infection control when handling all types of biospecimens. The CDC has offered some safe-handling guidelines for laboratories and related facilities. These guidelines include:

·   Laboratories should perform a site-specific and activity-specific risk assessment and continue to follow standard precautions when handling any biospecimen.

·   Environmental Protection Agency-approved disinfectants should be used to control viral pathogens.

·   Biospecimens which are suspected to be positive or confirmed SARS-CoV-2 positive specimens, isolates, or cultures, should be packaged and shipped as a Category B UN 3373 Biological Substance.

General Biospecimen Handling

Biospecimen quality has a direct and significant impact on diagnostic testing, clinical and preclinical research. Specimens of poor quality can lead to inaccurate test results and experimental errors. Because we understand the importance of high-quality specimens, our biorepository handles all specimens with care and precision, so our clients receive the highest quality of specimens. We observe the strictest guidelines including those set forth by the CDC and WHO to maintain sample quality across the board.

Labs must be adamant about ensuring COVID-19 lab safety and following standard biological safety procedures whether handling COVID-19 and other biospecimens. Staff should have PPE (personal protective equipment) available for staff including disposable gloves, eye protection, and lab coats or gowns. Biobanks should have a dedicated area for staff to put on or remove PPE.

The World Health Organization and the Centers for Disease Control and Prevention have both stressed the importance of using standard precautions. These precautions include decontaminating workspaces, treating waste to ensure viral particles are inactive, and eliminating unguarded aerosol production. As the pandemic continues to unfold guidelines will continue to be updated.

WHO and CDC Biosafety Guidance During the Covid-19 Pandemic

The CDC and WHO have issued interim biosafety guidance. Here are some of the current recommendations for the handling of biospecimens if they have known or are suspected to have contacted COVID-19.

·   Continue strict observance of relevant protocols including having all risk assessments and procedures conducted by capable personnel.

·   All lab techs should continue to wear appropriate PPE.

·   Processing should be conducted in a validated biosafety cabinet or a primary containment device.

·   The level of biosafety will depend on the nature of the work. BSL-2 for non-propagative work, and BSL-3 for propagative work.

·   All procedures in the bio lab should be done in such a way as to minimize the generation of aerosols.

·   Proper handling of biolaboratory waste.

·   Biospecimens should be placed in a secondary container if they are being transported.

·   Proper shipping of biospecimens based on their substance category.

All biospecimens should be handled as if they are infectious eliminates switching between protocols. Handling them as though they are infections also minimizes their exposure potential and the types of implications such exposure could have on the community and lab personnel.

There are exceptions when handling nucleic acid extracts, formalin-fixed, and paraffin-embedded (FFPE) tissue. When these processed biospecimens are handled using standard precautions and wearing appropriate PPE, no special considerations are necessary. Based on earlier studies of coronaviruses, the routine histologic fixation and processes should render SARS-CoV-2 inactive.

You can be assured that the products received from our repository are handled according to the ethical standards set forth by the World Health Organization. 100% of collected tissue samples are subjected to our strict quality control and all biospecimens are collected in accordance with our strict Standard Operating Procedure.

Resources:

https://academic.oup.com/ajcp/article/155/1/55/5911469

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7543273/

https://www.biobanking.com/safety-for-biobanks-strategies-for-safe-covid-19-specimens-handling/

https://research.ucsf.edu/covid-19-biospecimen-guidelines

Interim Guidelines for Biosafety and COVID-19 | CDC

What is Pathology and What Does a Pathologist Do?

A pathologist is a medical professional who examines bodies and body tissue. They are also responsible for conducting lab tests. A clinical pathologist is an essential part of medical teams who reach diagnoses for patients. After completing medical school, an individual must also complete three years of advanced medical education in a residency program before they are eligible to take board certification exams. Most pathologists are trained in both anatomical and clinical pathology.

The Role of Blood and Pathology Tests

Blood and pathology tests are essential for detecting, diagnosing, and treating disease. The term, “pathology” means the study of disease as well as causes and progression. There are a variety of pathology tests including blood tests, urine, stool (feces), and body tissue testing. Pathologists interpret the results of blood and pathology tests. They are looking for abnormalities in the samples that may indicate the presence of disease or health risks like cancer, chronic illnesses, or pre-diabetes. 

Why Blood and Pathology Tests Are Ordered

While pathology plays a role in detecting and diagnosing diseases, the tests are also important for other reasons including:

·   Properly treating a disease

·   Monitoring the progression of disease

·   Preventing disease

·   Determining the future risk of disease

·   Aiding research for new treatment options

·   Ensure the safety of treatment and procedures

When a doctor or specialist requests pathology tests it’s usually due to a concern about your health risks. A pathology test is effective for discovering if a problem or concern exists.

Types of Tissue Pathologists Examine

A pathologist is trained to examine tissue samples including samples as small as a dozen cells. Tissue cells may be obtained by aspiration or a needle biopsy. Larger tissue samples are surgically removed.

What do clinical pathologists do?

A clinical pathologist examines blood, urine, and other types of bodily fluids under a microscope. They are watching for the presence of certain chemicals or other substances. Their test results often determine a diagnosis or treatment option. Specimens used in clinical pathology include:

·   Blood Samples are used in many tests and can be checked in a variety of ways whole, plasma (the remaining fluid after red and white blood cells are removed), or serum (the clear fluid that separates from the blood during clotting).

·   Urine Samples are collected in a variety of ways including catheterization, clean catch specimen, or randomly.

·   Sputum Samples or phlegm samples are coughed into a clean container.

·   Stool Samples are often examined for the presence of blood.

·   Other Bodily Fluid Samples may include spinal fluid, pleural fluids, belly fluids, joint fluids, or bone marrow.

Clinical pathologists are often responsible for blood banks at hospitals. Their duties include collecting and processing blood products. They may also look at transfusion reactions or check tissue compatibility for transplants.

Conclusion

Pathology is a medical field that is quickly becoming more specialized. Pathologists provide experience and expertise when it comes to interpreting laboratory test results and evaluating cells, tissue, and organs in order to diagnose disease. A pathologist may determine a specific type of cancer and what stage it is in so that appropriate treatment can be recommended. In a quickly advancing technological age, their work is far from done and their importance continues to become apparent.

https://www.mskcc.org/cancer-care/diagnosis-treatment/diagnosing/role-pathology

https://pathology.uic.edu/understanding-your-pathology-report/

https://www.rcpath.org/discover-pathology/what-is-pathology.html

https://www.urmc.rochester.edu/encyclopedia/content.aspx?contenttypeid=85&contentid=P00955

https://www.betterhealth.vic.gov.au/health/conditionsandtreatments/Blood-and-pathology-tests

https://www.hopkinsmedicine.org/health/treatment-tests-and-therapies/the-pathologist

5 Laws That Apply to Biorepositories

With the increase of biological and pharmaceutical research, biorepositories are surfacing as a huge, important component. Biorepositories store and catalog a variety of biological samples. They must adhere to stringent regulations when handling and using biological specimens, especially when human samples are involved. Here are some of the legal guidelines biorepository staff much be familiar with and adhere to.

HIPAA Compliance

HIPAA or the Health Insurance Portability and Accountability Act is a US law. It is designed to protect patient’s medical records and privacy. Any business that handles any type of patient information much follow HIPAA regulations and practices to protect patient privacy. Biorepositories store samples were taken from human patients. If there is a breach in patient data or if the biorepository is not compliant with the guidelines set forth in HIPAA, they can be subject to criminal or civil penalties.

Informed Consent and Human Tissue Samples

The IRB (Institutional Review Board) oversees studies at research institutions when they involve human subjects. The review board is set up to help protect people from possible negative consequences when they are involved in research studies. In most cases, specimens collected from research participants or patients require the signing of an informed consent form approved by the IRB. Specimens are often stored in biorepositories for future use. These specimens require signed informed consent documents. It is often difficult to know when specimens will be used in research, donors may be able to withdraw their authorization for tissue sample use.

Patient’s Right to Withdraw from Research

When patients consent to the use of their samples for future research, they retain their right to withdraw that consent. As far as sample management and protocol in general, it can vary between institutions. Biorepositories should outline their best practices and how the specimens will be handled in cases where a patient withdraws consent. For instance, biorepositories may choose to destroy the sample, or they may be able to transfer the sample to a different donor-approved research project. Having a transparent operating procedure to follow for cases where donors withdraw consent, helps protect biorepositories from unintentional violations of HIPAA or IRB regulations.

Custody and Ownership of Samples

The laws vary from state to state and between institutions as they pertain to biological repository sample ownership and custody. In most cases, collected samples are owned by the organization that gathered them. Researchers who collect samples usually retain custody of them since they are using them for a particular study or purpose. Other times, they are in the custody of an IRB-approved biospecimen regulatory body. Biorepositories should have clearly stated guidelines for the ownership, custody, and distribution of samples. They should also have a clear plan for cases when a researcher leaves their institution.

Access to Biological Specimens and Data

Biorepositories also need to tightly control access to biological samples and the data associated with them. This helps ensure both the integrity and security of the specimen. Access to samples should be limited to authorized researchers and specific biorepository staff. Logs should be diligently maintained by biorepository staff. These logs should contain detailed audit trails for each sample including access logs.

Biological research is growing rapidly and as with other technologies, the pace is often faster than regulations. Laws that affect biorepositories are still relatively young and it’s likely that they will change as products, research, and technology continue to advance. Staff members at biorepositories should continue to be aware of regulatory developments as they occur in the future.

Importance of Bone Marrow Samples

Inside some large bones is a soft, sponge-like tissue called bone marrow. Bone marrow is important to the body as it contains cells that make up the immune system as well as stem cells that produce blood cells. Stem cells in the bone marrow mature into a variety of types of cells, each of which has specific vital functions that benefit the body. There are two types of bone marrow – red bone marrow and yellow bone marrow.

Distinguishing the Two Types of Bone Marrow

Red Bone Marrow

Red blood cells are present in the medullary cavity of the body’s flat bones like the pelvic girdle and sternum. It contains hematopoietic stem cells which are responsible for forming blood cells. Hematopoietic stem cells can turn into three types of blood cells.  Each type has important functions that keep the body alive and healthy. The three types of blood cells formed in the stem cells found in the red bone marrow include:

·  Red blood cells responsible for transporting oxygen throughout the body.

·  White blood cells responsible for fighting off infection.

·  Platelets that help prevent excessive bleeding by aiding the blood in clotting following an injury.

Yellow Bone Marrow

Bone marrow also contains yellow bone marrow which is named because of its color caused by a high concentration of fat cells. Yellow bone marrow is surrounded by a layer of red bone marrow, and it can be found in the medullary cavity of long bones. The yellow bone marrow contains marrow stromal cells or mesenchymal stem cells. They help produce fat, bone, and cartilage. Yellow bone marrow is useful for helping store fat in cells called adipocytes. This action helps to maintain the right environment so that bones have the sustenance they need to function properly.

Function of Bone Marrow

Simply put, bone marrow’s function is to produce a variety of cells that are essential for sustaining life. Some conditions affect the health of the bone marrow. This can impair its ability to produce an adequate number of healthy cells. Bone marrow transplants administer healthy bone marrow through a central line for patients in need of them. There are several things a bone marrow transplant can help with including:

·  Healthy bone marrow can replace damaged or nonfunctioning bone marrow.

·  A bone marrow transplant can help restore the bone’s natural function after it’s been damaged due to treatments like radiation or chemotherapy.

·  It can prevent further damage caused by some genetic diseases.

·  Healthy bone marrow helps regenerate the immune system and helps fight off residual or existing cancers like Leukemia.

How Bone Marrow Samples are Used

One of the leading reasons for the need for frozen bone marrow samples is scientific research. There are numerous clinical trials being conducted around the world that are studying and assessing the benefits of using bone marrow for treating a variety of conditions and disorders including:

·  Cancers and tumors including lymphomas and Leukemias

·  Heart disease including CAD (coronary artery disease), heart failure, cardiomyopathies, and acute myocardial infarction

·  Organ failure including COPD, Crohn’s disease, diabetes, Cirrhosis, lupus, and chronic renal failure

·  Muscle-skeletal disorders including osteoarthritis, myopathies, scleroderma, and fractures

·  Peripheral vascular disease including poor circulation, gangrene, and non-healing wounds

You’ll find our biorepository carries a variety of frozen cells and bone marrow samples for use in single-cell analysis, genetics research, toxicology, treatment research, and much more.

What is Gene Therapy?

Genes within cells are critical to your health. If a gene is defective, it can make you sick. As medicine advances, researchers and medical professionals search for a way to treat defective genes, either by modifying them or replacing them. According to the Mayo Clinic, gene therapy is an experimental technique that replaces a faulty gene or adds a new, healthy gene to cure diseases or help your body fight diseases. 

Gene therapy has multiple applications, including: 

• Replacing mutated genes: Sometimes genes stop working correctly or don’t work at all. This can cause diseases that are difficult to fight, such as cancer. 

• Fixing mutated genes: Mutated genes can create diseases, or make it more difficult for your body to fight disease. 

• Making diseased cells easier to find in the immune system: Sometimes, your body can’t find an illness because your immune system doesn’t recognize it. In the future, gene therapy may help you train your immune system to recognize dangerous cells. 

Currently, gene therapy isn’t available as a medical procedure and is only available through clinical trials. As of 2017, The U.S. Food and Drug Administration has approved three forms of gene therapy products. Two of these products reprogram a patient’s existing cells to attack deadly cancer, and recently, the newest product targets a form of an inherited vision loss caused by mutation of a certain gene. Gene therapy will be beneficial in aiding in the treatment of cystic fibrosis, heart disease, diabetes, hemophilia, AIDS, cancer, and much more

Function of Cells and Genes

The FDA defines cells as the basic building blocks of living things. Human bodies are filled with trillions of cells, and within each cell, are thousands of genes. These genes provide the information needed to create muscles, bones, features, and more. Genes aid in the function of digestion, creating energy and physical growth. 

Genes also contain our DNA, which controls how our body forms. When a gene doesn’t work properly, our bodies might experience deformity or disease. Gene therapy attempts to address the genes that aren’t working properly.

How Does Gene Therapy Work? 

Sometimes a gene can be defective or missing at birth, or a gene can mutate in adulthood, causing health problems and diseases. Throughout the many clinical trials and research conducted on gene therapy, researchers have found that successful gene-based therapies rely on three elements, including: 

1. A well-defined disease gene

2. A therapeutic gene

3. An efficient gene delivery system, known as a vector

Gene therapy seems to be specifically successful in dermatology. As the largest organ on the human body, skin is easily accessible for gene delivery. It’s also easily manipulated and evaluated, making it a primary target for gene therapy. Having an efficient vector is required for successful gene therapy. These vectors are genetically engineered to deliver the gene to the host. Viruses are particularly good at delivering genetic materials into a cell, and can therefore be used as successful vectors. 

Gene Transfer Techniques

The vector will be injected either inside the body or outside the body, depending on which part of the body needs the gene therapy. The vector will contain the desired, healthy gene and is introduced to the cells. 

There are two techniques for gene transfer, including in vivo gene therapy and ex-vivo gene therapy: 

• In vivo gene therapy: Genetic material is transferred using a vector that is directly injected into the patient

• Ex vivo gene therapy: Cells are collected from the patient, genetically manipulated, and then administered to the patient

As we previously stated, viruses are particularly successful acting as vectors. Viruses are able to recognize specific cells and carry genetic material to the cell’s genes. Before the virus vector is administered to the patient, researchers will remove the disease-causing gene from the virus, and replace it with the desired genes required to stop the disease. Non-viral methods are possible and can be applied through electrical field-mediated gene transfer, magneto-permeabilization, and micro-needles.

Risks

Due to the method of utilizing viruses as a vector, there can be possible risks that it’s important to be aware of. 

The viral genetic application can present the following risks: 

• Unwanted immune system reaction

• Targeting the wrong cells

• Infection caused by the virus

• Possibility of causing a tumor

Our biorespository services offer tissues and cells from multiple collection samples, including melanoma, stomach tissue, lung tissue, kidney tissue, prostate tissue, and more. If you have any questions, contact our team.

Resources:

• https://premier-research.com/bylined-articles/gene-therapy-in-dermatology-transfer-techniques-and-delivery-systems/

• https://www.mayoclinic.org/tests-procedures/gene-therapy/about/pac-20384619

• https://premier-research.com/the-state-of-gene-therapy-in-dermatology/

• https://www.fda.gov/news-events/press-announcements/fda-approves-novel-gene-therapy-treat-patients-rare-form-inherited-vision-loss

• https://www.fda.gov/consumers/consumer-updates/what-gene-therapy-how-does-it-work

Human Artificial Chromosomes

Artificial chromosomes are constructed in the laboratory that contain DNA sequences and perform the primary functions of natural chromosomes. Artificial chromosomes are used for many purposes, such as introducing and controlling new DNA into a cell, studying how chromosomes function, and mapping genes in genomes. Here, we’ll analyze how natural chromosomes function and how artificial chromosomes are created and utilized. 

Natural Chromosome Function

Chromosomes are complex structures that dictate how genes can be translated into proteins and how existing genes are translated. According to the Biology Dictionary, “A chromosome is a string of DNA wrapped around associated proteins that give the connected nucleic acid bases a structure.” 

Gene Expression

The process of translating genes is known as gene expression, which is responsible for creating organisms. Cellular molecules regulate genes and transcription, and do so by engaging or disengaging proteins. 

Cell Division

Cell division occurs when a cell's DNA is distributed to daughter cells. There are two types of cell division in humans, including mitosis and meiosis. Mitosis is the process by which the cells produced by division have the same exact genetic information as the original cell. Meiosis is when the cells produced by division have only half the genetic information as the original cell. 

Artificial Chromosome Function

Artificial chromosomes may be a promising tool in genetic therapy applications and have many advantages over the current system in place. Artificial chromosomes can replicate and segregate on their own, without integration into the host chromosome. 

Mammalian Artificial Chromosome

Mammalian artificial chromosomes contain mammalian or human chromosomes. They are composed of repeated DNA sequences found at the natural centromeres of human chromosomes, which are referred to as alphoid DNA. 

Mammalian artificial chromosomes are used as vectors for delivering large fragments of DNA to mammalian cells, and to whole animals for the expression of large genes. Despite their potential, they have not yet been used in practical applications. 

Yeast Artificial Chromosomes

Yeast artificial chromosomes are used in yeast and mammalian cells to replicate the components of a single DNA molecule. According to Encylopedia.com, “In baker's yeast, telomeres, centromeres, and origins of replication have all been defined using genetics and have been cloned”. When these replications are assembled, they can grow into a small chromosome when placed in bacteria. This forms a vector capable of including up to a million bases of other DNA as a chromosome. 

This can be utilized to analyze the properties of yeast chromosomes but has also been used in the early phases of genome mapping projects. They can also be used to find information on the inheritance of genetic diseases.

Resources: 

• https://www.encyclopedia.com/science-and-technology/biology-and-genetics/genetics-and-genetic-engineering/artificial-chromosome

• https://www.nature.com/articles/gt2009102

• https://biologydictionary.net/chromosome/

• https://www.the-scientist.com/modus-operandi/streamlined-artificial-chromosome-creation-66612

Artificial Cloning: The Future of Medicine?

According to the National Human Genome Research Institute, “gene cloning produces copies of genes or segments of DNA”. Researchers have successfully cloned a range of biological materials such as genes, cells, tissue, and even sheep. Natural cloning exists, such as identical twins and in some plants or single-cell organisms such as bacteria. 

There are three types of artificial cloning done by researchers, including gene cloning, reproductive cloning, and therapeutic cloning. We’ll discuss how these various methods are conducted, their benefits, and more. 

Types of Artificial Cloning

Genes are cloned through a complex process conducted by researchers that insert a gene collected from an organism, often referred to as the “foreign DNA”, and inserted into the genetic material of a carrier, often referred to as a vector. A vector can be numerous things, including viruses, yeast cells, bacteria, plasmids, and more. Once the foreign DNA is inserted into the vector, it is placed in a laboratory environment that encourages it to multiply. 

Gene Cloning

Gene cloning (also known as DNA cloning) creates copies of genes or segments of DNA. 

This can be used for:

• Biopharmaceuticals: the creation of insulin or human growth hormones

• Gene therapy: When patients lack the functional form of a specific gene 

• Gene analysis: to build an artificial, mutated version of a gene to help them understand how normal genes are supposed to function and how they may mutate 

Gene cloning is a complicated and thorough process, but it can be incredibly beneficial to healthcare professionals and the future of medicine. DNA cloning allows healthcare professionals to analyze the function of a gene, investigate a gene’s characteristics, potential mutations, 

Therapeutic Cloning

Therapeutic cloning is the process of creating a cloned embryo for the purpose of creating embryonic stem cells with the same DNA as the donor. There have not been any human embryos created through therapeutic cloning so far, but might be a possibility once the resources and technology are available. Therapeutic cloning can grow healthy tissues in the laboratory to replace injured or diseased tissue, making this process particularly beneficial for cancer patients. It has the potential to create organs (allowing patients to get off the ever-growing donor list), treat tissue rejection, and can act as a preventative treatment.

Reproductive Cloning

Reproductive cloning is the process of producing copies of an entire animal. While the idea of DNA cloning is not new, the first cloned animal was Dolly the sheep in 1996. Collected DNA that is intended to be replicated is transferred to the donor animal’s somatic cell into an egg cell and is then developed into an early-stage embryo in a test tube. These test tubes are designed to replicate the womb of an adult female animal.

Artificial cloning is a complex process and raises many questions about medical ethics. However, we cannot deny the overwhelming benefits it can provide. Using DNA cloning to gain a better understanding of genetic mutations provides researchers with endless opportunities to explore how genes and DNA function. The use of therapeutic cloning will allow us to provide sick patients with alternative choices. We are discovering more about the benefits and the risks of genetic cloning, so it is important to stay updated on the latest findings. Explore our biorepository to order a variety of cell and tissue samples for your own research and medical use

Resources:

• https://www.genome.gov/about-genomics/fact-sheets/Cloning-Fact-Sheet

• https://geneticeducation.co.in/gene-cloning-definitions-steps-procedure-applications-and-limitations/

• https://www.khanacademy.org/science/ap-biology/gene-expression-and-regulation/biotechnology/a/overview-dna-cloning

What is Direct-to-Consumer Genetic Testing?

Genetic testing is the process of analyzing DNA to identify changes or permutations in gene sequence or expression. Genetic testing helps physicians in the process of diagnosis for illnesses such as cancer, chronic illness, or chromosomal disorders such as Autism or Cerebral Palsy. 

Genetic testing is performed for countless reasons, including: 

• Prenatal screening and diagnosis to identify any present defects the unborn child may have

• Preimplantation genetic diagnosis examines embryos during in vitro fertilization

• Carrier screening to identify if a person has a gene or genetic disorder without showing any symptoms

• Forensic testing to identify and analyze criminal evidence 

• Paternal testing to identify the father of a child

• Presymptomatic testing to identify if an asymptomatic person at risk of a condition has developed the disease-causing mutation. 

• Presymptomatic testing to identify the risk of developing a disease 

The process of genetic testing is thorough and often includes a biopsy (the collection of tissue or genetic sample), analysis of the collected tissue sample, and proper storing of the tissue sample. Healthcare professionals are very thorough with this process so they can accurately identify and diagnose a condition. 

However, genetic testing can serve many more purposes outside of physician diagnosis, such as Direct-to-Consumer (DTC) genetic testing. Direct-to-Consumer (DTC) genetic testing is sold directly to consumers without requiring the assistance of physicians or other healthcare professionals. Many people are interested in DTC genetic tests to obtain and analyze information on their ancestral background, medical history, and much more. 

Risks and Benefits of Direct-to-Consumer Genetic Testing

DTC testing is marketed to individuals through television, advertisements, the Internet, radio, and more. Individuals will send a sample of their DNA to obtain results through an online or written report. These tests are popularly used for ancestral information, common traits in their genetic lineage, predictions about health, and more. All of this can be done without the assistance of a healthcare provider, which is what attracts so many customers. Just like all healthcare testing, there are benefits and risks to DTC as well. 

Benefits

The primary benefit of DTC testing is that it brings awareness to genetic and health conditions and helps customers understand the importance of this awareness. These tests are easily accessible to people who may not have access to a clinician for contemporary genetic testing. These samples are often obtained through non-invasive means, such as providing a swab of saliva, rather than through a biopsy that can be intrusive and complicated. Additionally, these tests are often less expensive than traditional genetic testing. 

Risks

There are currently no regulations on DTC genetic testing, so it is important to do thorough research before ordering a test. While these tests provide generic information on health conditions, it does not provide conclusive results. If you believe you have a serious condition, such as cancer, it is important to seek professional medical assistance. Engage in DTC genetic testing with caution.

In conclusion, DTC genetic testing can be beneficial in providing information on genetic profiles for a low cost in a more convenient way than traditional testing. However, it is important to be aware of their shortcomings. Discuss any concerns you may have with your medical professional. 

Resources:

• https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6808639/

• https://medlineplus.gov/genetics/understanding/dtcgenetictesting/directtoconsumer/

• https://medlineplus.gov/genetics/understanding/dtcgenetictesting/dtcrisksbenefits/

• https://medlineplus.gov/genetics/understanding/dtcgenetictesting/dtcrisksbenefits/

How to Diagnose Chromosome Abnormalities

Our chromosomes are essentially the blueprint of our body. Consisting of proteins and DNA, they carry the genetic information of our body. Humans have 23 pairs of chromosomes. 1 through 22 pairs of numbered chromosomes are called autosomes, while one pair is a sex chromosome, either X and Y. A cytogeneticist is a medical professional that is dedicated to examining a human's chromosomes under a microscope. They will identify how the chromosomes are arranged, which will help lead to a diagnosis of the patient. 

There are multiple approaches to analyzing discrepancies, including the molecular approach and the cytogenetic approach. 

Cytogenetic Approach

The cytogenetic approach identifies chromosome abnormalities. After chromosomes are fixed and sit in cell culture before they spread on microscope slides and stained. The staining methods help distinguish between each individual chromosome and are analyzed under a microscope or in pictures.

Molecular Approach

Microarray analysis is utilized for the molecular approach. A microarray is a method used to detect the expression of thousands of genes at once. Microarray slides have thousands of tiny spots that consist of a DNA sequence of genes, often known as a gene chip or DNA chip. You can learn more about our microarray process here.     

Prenatal Chromosome Testing

Prenatal chromosome testing will help you and your medical professional determine if further diagnostic testing is required. If the screening test indicates that there is a possibility that your baby has Down syndrome, trisomy 13 or trisomy 18, or other chromosome abnormalities. These tests are able to diagnose 99.9% of chromosomal abnormalities. 

• First trimester screen: The first trimester screening utilizes blood work and an ultrasound to measure the fetal nuchal translucency. According to Beth Israel Deaconess Medical Center, this test detects 85% of fetuses with Down syndrome and 90% of fetuses with trisomy 13 or trisomy 18.

• Quadruple screen: A maternal serum quadruple screen, also known as a second-trimester screen, is conducted by taking a blood test between 15 to 20 weeks gestation. This test provides an estimation of the chance that a pregnancy will be affected by chromosome abnormalities. This test detects 80% of fetuses with Down syndrome, 65% of fetuses with trisomy 18, and 90% of fetuses with open NTDs. 

• Sequential screen: A sequential screen utilizes both the first and second-trimester screens that increase the detection rate of Down syndrome and trisomy 18. Once you receive the results for your first-trimester screen, you will have the option of getting your second-trimester screening. If you decide to get your second-trimester screening, the results will incorporate the results from both screenings. Sequential screening detects more than 92% of fetuses with Down syndrome and greater than 92% of fetuses with trisomy 18. 

Our biorepository offers a wide variety of testing including microarrays, frozen tissue samples, biofluid samples, blood samples, and more. Contact us to learn more today!  

Resources:

• https://www.genome.gov/genetics-glossary/Chromosome

• https://www.chromosome18.org/diagnosis-of-chromosome-abnormalities/

• https://fenwayhealth.org/wp-content/uploads/Chromosomal-Abnormalities-Brochure.pdf

• https://www.nature.com/scitable/definition/microarray-202/

How Breast Cancer Tissue is Collected

Most people discover breast cancer by noticing a mass or abnormal feature on their chest during self-examination by themselves or with their doctor. Sometimes, these abnormalities are discovered during a screening mammogram, and in some instances, women will notice a swollen breast or red nodule under their arm. The process of diagnosing and testing cancer cells can be an elaborate process. There’s a wide variety of tests to pick from, based on your age, health, symptoms, and the results of earlier medical tests. 

These tests include, but certainly not limited to, the following: 

• Imagining Tests: Diagnostic mammography, ultrasound, and MRI

• Biopsy: Fine needle aspiration, core needle, surgical, image-guided, and sentinel lymph node.

Doctors and pathologists will conduct many tests in the process of diagnosing and treating breast cancer. They’ll decide which tests are the best option based on your age and overall health, what symptoms you’re showing, results of prior medical tests, and the type of suspected cancer. 

Imaging Tests for Breast Cancer

Imaging tests are conducted to show pictures of what’s inside your body. These are beneficial to see the areas surrounding the cancerous tissue, how far cancer has spread, and if the treatment is working. 

MRI

MRI’s is a popular option because it’s painless and doesn’t require any special preparation, as well as providing a detailed image of what’s happening in your body. An MRI uses magnetic fields while a special dye is given to the patient before the screening. This dye helps the image be as clear as possible. Doctors may routinely schedule MRI tests during treatment in order to surveil the cancerous tissue and adjust the treatment as needed. 

Ultrasound

Ultrasounds are useful for tracking changes in breast tissue, especially changes that can be felt but don’t appear on a mammogram. Ultrasounds are helpful in differentiating fluid-filled cysts and solid masses, which can help them determine if further testing for cancerous cells is necessary. They’re a popular testing choice for many practitioners and can be conducted safely, as they do not expose the patient to radiation. 

Diagnostic Mammography 

Diagnostic mammography is often used when a woman is experiencing symptoms of breast cancer, such as an unexplained lump or discharge from the nipple. It may also be used to get a more detailed image if something suspicious appears on a screening mammogram. 

Biopsy for Breast Cancer 

There are numerous variations of biopsies for cancer cell collection. While imagining tests can help detect the presence of cancerous tissue, biopsies are the only way to get a definite diagnosis of cancer. Different biopsies are classified by the technique used to collect the tissue and the size of the needle. Because there are so many biopsy techniques, we’ll go over the most popular options for breast cancer patients according to the American Cancer Society. 

Fine Needle Aspiration

An FNA biopsy (fine needle aspiration), a very thin and hollow needle is attached to a syringe a very small amount of tissue from the suspected area. This needle is even thinner than the size used for blood tests, so it’s minimally invasive. 

Lymph Node Biopsy 

In some instances, a doctor may need to conduct a biopsy on the lymph nodes underneath the arms to see if cancer has spread. This can be conducted in multiple ways, either during surgery when the breast tumor is removed, with a sentinel lymph node biopsy, or an axillary lymph node dissection. Your doctor will know the best option based on your needs. 

Surgical Biopsy

Sometimes surgery is required to remove the specimen needed to properly test the potentially cancerous tissue. During these procedures, the doctor or surgeon will typically remove the entire mass or abnormal growth as well as the surrounding breast tissue. 

Core Needle Biopsy

A core needle biopsy is similar to fine-needle aspiration except it uses a larger needle. These are common when a doctor has felt changes or seen suspicious areas on an ultrasound, mammogram, or MRI. This is typically the preferred type of biopsy for breast cancer patients. 

How are Breast Cancer Tissues Analyzed? 

After the tissue is collected from a biopsy, the sample is sent to a lab where a pathologist will analyze it and provide the doctor with their findings for an official diagnosis. Your medical professionals will consider multiple factors while analyzing the cancerous tissue. 

Tumor Features 

Pathologists will study the tumor under a microscope to determine if the cancer is invasive or non-invasive, ductal, lobular, or another type of cancer. They’ll also determine if cancer has spread to the lymph nodes by measuring the distance from the tumor to the edge of the tissue removed. This is known as the margin width. 

ER-positive and PR-positive Cancers 

ER and PR analysis determine the patient’s risk of cancer returning and what kind of treatment will decrease the chance of reoccurrence. Hormonal therapy is usually used to decrease the chance of the cancer returning. 

Grade 

The grade of a tumor refers to the differentiation between cancerous cells and healthy cells and how fast or slow they are growing. “Well-differentiated” cancer cells look similar to healthy tissue and have different cell groupings, while “poorly-differentiated” cancer cells look very different from healthy tissue. Grade is categorized by three levels including grade 1 (well-differentiated), grade 2 (moderately differentiated), and grade 3 (poorly differentiated). 

Summary

It’s important to conduct regular self-examinations to track any abnormalities that appear on your breast, or you can ask your doctor to regularly conduct examinations. Doctors and pathologists have endless options available to them and will use the tests that are best for you and your situation. Our biorepository collects and safely store cancerous cells, utilizing tissue microarrays, frozen tissue samples, biofluid samples, and more. 

Contact us today to learn more. 

Sources:

• https://www.cancer.org/cancer/breast-cancer/screening-tests-and-early-detection/breast-ultrasound.html

• https://www.cancer.org/cancer/breast-cancer/screening-tests-and-early-detection/breast-biopsy.html#:~:text=A%20core%20biopsy%20uses%20a,if%20breast%20cancer%20is%20suspected.

• https://www.cancer.net/cancer-types/breast-cancer/diagnosis

• https://www.hopkinsmedicine.org/health/treatment-tests-and-therapies/mammogram-procedure

How Does Germ Immunization Work?

Germs are all around us and impossible to avoid. There are many precautionary measures we can take to protect our bodies from germs, and our body even has natural defenses such as mucus and microscopic hairs, but sometimes, we need help to fight harmful pathogens (organisms that cause disease). When a pathogen is introduced to the body, the body responds by triggering its immune system. 

The Immune System 

When harmful germs such as bacteria or viruses enter the body, they attack and multiply, leading to an infection and then illness. The National Cancer Institute defines the immune system as a complex network of cells, tissues, organs, and substances that assist the body in fighting infections and disease. This complex system contains white blood cells, the spleen, tonsils, lymph nodes, bone marrow, and much more.

White blood cells consist of macrophages, B-lymphocytes, and T-lymphocytes.

• T-lymphocytes: A defensive white blood cell that attacks infected cells in the body.

• B-lymphocytes: A defense white blood cell that generates antibodies to attack the antigens left behind by macrophages.

• Macrophages: White blood cells that swallow and digest germs, dead cells, or dying cells. They leave behind parts of the invading germs called antigens, which are identified as dangerous and stimulates the antibodies to attack them.

It takes the body several days to produce and utilize all the germ-fighting tools to fight the infection or illness. After the infection, however, the immune system utilizes T-lymphocytes, also known as memory cells, to teach the body how to respond if the same illness is encountered again.

T-lymphocytes attacking a virus

How do Vaccines Work?

Vaccines develop immunity against an illness by imitating the infection, causing the immune system to produce T-lymphocytes and antibodies. This treatment rarely causes illness, but a few side effects are expected, which can be discussed with health professionals. Once the imitated infection is no longer present in the body, the immune system is equipped with the necessary memory cells to properly fight the illness if encountered in the future. It typically takes a few weeks for the body to produce the necessary T-lymphocytes and B-lymphocytes.

Types of Vaccines

There are five primary types of vaccines that are utilized in the U.S. These practices differ from region to region around the globe, depending on their resources and needs.

• Live, attenuated vaccines: These vaccinations are designed to fight viruses and bacteria. They contain a version of the living virus or bacteria. However, this duplicate illness is weakened so that it does not cause harm to the vaccine recipient. These vaccines are great teachers of the immune system and are very effective in teaching the body how to fight off specific diseases.

• Inactive vaccines: These vaccines also fight viruses and bacteria and are created by inactivating the germ during the process of making the vaccine. Inactivated vaccines produce immune responses differently than live vaccines, and oftentimes require multiple doses to build up and maintain immunity.

• Toxoid vaccines: These vaccines prevent diseases caused by bacteria that produce toxins in the body. Similar to other vaccines, the toxins are weakened in the process of creating the vaccine. When the immune system is introduced to a toxoid, it’s able to fight natural toxins in the future.

• Subunit vaccines: Instead of containing the entire pathogen like other vaccines, the subunit vaccine only includes the antigens that stimulate the immune system. This vaccine is designed to be safer and easier to create. This means side effects are less common.

• Conjugate vaccine: This type of vaccine combines a weak antigen and a strong antigen. Different kinds of bacteria have antigens with an outer coating of sugar-like substances called polysaccharides. This sugar-like coating makes it more difficult for immature immune systems to recognize it and properly respond. This method helps the immune system create a stronger response to the weak antigen.

Many people, especially babies and teenagers, may need more than one dose when receiving a vaccine for the first time. It’s common for the initial exposure to a vaccine will not be sufficient to build complete and efficient immunity.

Herd Immunity

Ever wondered why some vaccines are required for school or other social activities? Vaccines don’t just work on the individual level - they are also responsible for protecting an entire population. If one person was vaccinated against a dangerous illness, it wouldn’t be beneficial if the rest of the population wasn’t vaccinated for the same thing. Once enough of the population is vaccinated, the possibility of an outbreak becomes less likely. The bacteria or virus has a difficult time finding enough eligible hosts, so they eventually die out entirely. This process is known as herd immunity and is critical in protecting ourselves and the people around us.

Because new viruses, bacterias, variants, and mutations of viruses are always present and ever-changing, vaccine research is always developing as well. Our biorepository services provide various tissues from various donors. Contact us today to learn more.

Resources:

• https://vaccineinformation.org/how-vaccines-work/

• https://www.niaid.nih.gov/research/vaccine-types

• https://www.publichealth.org/public-awareness/understanding-vaccines/vaccines-work/

• https://www.cdc.gov/vaccines/hcp/conversations/downloads/vacsafe-understand-color-office.pdf

Tissue Microarray Method (TMA) for Bone Marrow Biopsies

Tissue microarray (TMA) is a method in pathology that allows researchers to analyze a greater number of tissues than traditional biopsies. Researchers can analyze and test hundreds of tissue samples with customized antibodies for each all at once. This makes tissue microarrays helpful for saving and optimizing time and resources. Here, we’ll discuss the findings of research conducted to apply TMA methods to bone marrow biopsies, the use of TMA for bone marrow biopsies, and the benefits and drawbacks of this new research method.

What is a Tissue Microarray?

TMA’s are typically used to analyze and diagnose cancerous tissue cells. Small tissue samples are collected, embedded with paraffin, and arranged in a paraffin block. A microarray block can include up to 100-500 sections. Each sample can be injected with its antibody, allowing researchers to analyze various samples at once without using excess resources.

Benefits of Tissue Microarray

Paraffin blocks preserve the tissue samples, allowing them to last much longer than other tissue sampling methods. This allows researchers to revisit the samples if need-be and allows them to analyze how they change over an extended period. Tissue microarray is cost-efficient and time-efficient, making it the future of diagnostics and research in the field of medical science and hematopathology (the study of disease and disorders).

Bone Marrow Biopsy (BMB)

Bone marrow biopsies are used to analyze patients who may have trouble creating blood cells. The process of the biopsy involves removing a small amount of bone marrow that exists within your bones. Bone marrow is a soft tissue that is created in the center of large bones, and samples are typically collected from the front of the pelvis.

• A pathologist may recommend a bone marrow biopsy if you experience the following:

• Anemia (lack of red blood cells)

• An abnormal amount of blood cells

• Iron deficiency

• Cancer present in blood-forming tissue such as leukemia or lymphoma

• Cancer that has spread to the bone marrow

• Symptoms of chemotherapy

Drawbacks of Bone Marrow Biopsy

Most bone marrow biopsy samples are very small in size due to how invasive they are. This provides health professionals and researchers with a smaller sample to study. A huge advantage of TMA’s is that they provide researchers with sufficient tissue samples. Most bone marrow biopsies are typically no larger than 2.0mm wide and 2.5 cm long. TMA methods, however, require tissue samples of 3-4mm.

According to research conducted by Obermann, Marienhagen, Stoehr, Wuensch, and Hofstaedter, “Availability of TMAs from bone marrow biopsies (BMBs) would be especially useful in hematopathology research because a large number of neoplasms of the lymphatic and hematopoietic system involve the bone marrow. In some entities, such as leukemia, BMBs can be the only type of tissue containing neoplastic cells that is available for the histopathologist.” Therefore, the cost-efficiency and time-efficiency of TMA methods would be incredibly useful for bone marrow biopsies.

Tissue Microarray constructed from Bone Marrow Biopsies

TMA for Bone Marrow Biopsy Utilizing the Stacking Method

The research previously mentioned adapted tissue microarray methods to better suit bone marrow biopsy samples and meet the specific requirements of BMB. Bone marrow biopsies were selected and inserted into pre-made holes in the TMA block, with a thickness of 0.1 mm and a diameter of 1.6mm. The researchers found that this size optimized the preservation of the biopsy, which is a primary desired advantage of the TMA method.

The Stack Method

The stack method was then used by stacking three punch bone marrow biopsies from one donor block on top of another. The stack method resulted in the final size of the tissue block being 4mm, which is the size required for efficient TMA testing.

Conclusion

After successfully constructing a TMA for a bone marrow biopsy, a total of 100 cells were assessed both in whole sections and tumor cores. The TMA contained 33 cores of 11 BMB’s, and the bone marrow was well preserved - achieving the study’s goal. Tissue microarray is an advantageous method and the possibilities of its application should and can be explored to further our ability to efficiently research, analyze, and diagnose.

Explore our biorepository services and the various cancer types we collect for tissue microarrays.

Resources:

• https://www.future-science.com/doi/10.2144/000112073

• https://www.jpatholtm.org/journal/Figure.php?xn=kjpathol-46-562.xml&id=

• https://en.wikipedia.org/wiki/Tissue_microarray

• Limberger, K.A., Bogatyreva, L., Todorova, R. et al. Tissue microarray technique is applicable to bone marrow biopsies of myeloproliferative neoplasms. Histochem Cell Biol 147, 75–82 (2017). https://doi.org/10.1007/s00418-016-1476-x

Blood Donation for Cancer Patients

Blood is an essential part of our body’s mechanism that is necessary for us to function. Blood helps transport nutrients such as oxygen into the lungs and tissues, prevent excessive bleeding by creating blood clots, and carry cells and antibodies to fight disease or infection. Cancer patients are particularly in need of blood transfusions. Here we will discuss the various ways you can help those suffering from cancer, and how a biorepository clinic can help.

Blood is made up of numerous cells that help it function properly. These cells include:

• Red blood cells: Red blood cells are critical to fighting disease. They contain hemoglobin, a protein molecule that plays a critical role in maintaining the shape of red blood cells. It is responsible for transporting oxygen and carbon dioxide throughout our blood. The iron within hemoglobin also causes the red color you see in blood. Hemoglobin levels can be tested by machines designed to study blood. The normal hemoglobin range is 4.5-6.2 million for men and 4.0-5.2 million for women.

• Platelets: Platelets are a small, colorless disk-shaped cell fragment that is responsible for the self-healing functions of our body. They are found in large amounts within the blood and help create blood clots to prevent bleeding. Healthy platelets assist in the process of hemostasis, thrombosis, and wound healing. When platelets are low, life-threatening bleeding can occur, which is particularly dangerous for cancer patients who might have frequent surgeries or treatments.

• Plasma: Plasma is a yellow liquid that carries the red blood cells, platelets, cells, proteins, and antibodies. Plasma donations are particularly important because they contain a very high concentration of blood-clotting proteins, however, not many cancer patients require plasma transfusions.

Why Blood Donations are Necessary for Cancer Patients

Cancer is incredibly taxing on the human body and requires a lot of assistance from medicine and blood products. Extra blood is crucial to their treatment, so healthy individuals need to donate blood when possible. Some patients will require extra whole blood or small portions known as platelets. Cancer or the treatment of cancer can cause low red blood count in patients, which is necessary for them to effectively fight their disease.

Here are two common symptoms cancer patients experience during treatment, and how blood transfusions can help:

• Anemia: Anemia occurs when you lack enough healthy red blood cells to transport enough oxygen to the tissue in your body, leaving you feeling fatigued and weak.

• Thrombocytopenia: Thrombocytopenia occurs when the bone marrow in a patient’s body is damaged. This damage can occur through chemotherapy, a common treatment for many cancers such as lymphoma or leukemia.

Many blood donation organizations will check donated blood for unhealthy red blood cell antibodies that might harm the receiving patient, and the healthy samples are stored in a blood bank.

What to Expect

While there is a high demand for blood donations, there are many factors that must be considered before accepting a donor. Here are some things you can expect the blood bank to consider before you become a donor:

• Medication: People who are prescribed or are taking blood-thinners (ex. Aspirin, Warfarin, Eliquis, Heparin, etc) may have to wait until the medication has been removed from their system. People who take antibiotics to fight infection will have to wait until they are healed and off antibiotics before they can donate.

• Certain Health Conditions: People who have been diagnosed with low blood pressure, heart conditions, or viruses such as HIV or hepatitis, typically cannot donate blood to cancer patients.

• Pregnancy: Pregnant women are not allowed to donate blood and must wait 6 weeks after giving birth to donate.

• Travel: Traveling to or living in certain countries can disqualify you from donating blood to cancer patients. The countries that may disqualify you typically have high rates of malaria or other viruses. Typically you can wait to donate, however.

• History of Cancer: People who have suffered from cancer, particularly leukemia or lymphoma, in the past may not qualify to donate blood to current patients.

Platelets Donation Process

Donating platelets is similar to donating blood, and the process is known as apheresis. Blood is removed from the arm and a centrifuge separates the platelets from the blood. The remaining blood is returned to the donor. This process maximizes the number of platelets collected, rather than a regular, whole-blood donation. People who are planning on donating platelets will be given instructions to prepare for the donation.

Plasma Donation Process

Plasma donation is known as plasmapheresis and requires a process that can take up to an hour. Donors are required to be at least 18 years of age, must weigh at least 110 lbs, and must complete two separate medical screenings. These medical screenings include a medical history screening and a test for transmissible viruses. If these requirements are met, an individual may donate plasma to cancer patients. Donating plasma is completely safe, however, your hemoglobin levels must be tested after to ensure your health.

Speak with your doctor before donating blood and they will be able to properly prepare you, or advise you against it if it’s not in the best interest of your well-being.

How a Biorepository Can Help

Biorepositories act as a library of biospecimens for medical research to further advance our treatments and medicine as a whole. If you don’t qualify to donate blood to cancer patients, you can still donate your blood to a biorepository for medical research and help scientists conduct research. We proudly collect whole blood samples, serum samples, and plasma samples. Our blood and plasma sample inventory is collected from a variety of donors, including:

• Systemic Autoimmune Condition (i.e., Lupus, rheumatoid arthritis, osteoarthritis, diabetes, psoriasis, atopic dermatitis, and more)

• Benign condition

• Cancer

• CNS Disorders (i.e., depression, bipolar disorder, autism, ADHD, addiction, and more)

• Normal donors

• Pediatric conditions

• Pregnant donors

If you are interested in purchasing any of our samples, feel free to contact us.

Resources

• https://www.medicinenet.com/hemoglobin/article.htm

• https://www.cancer.net/blog/2017-03/help-people-with-cancer-donate-blood-and-platelets

• https://www.drugwatch.com/health/cardiovascular-health/blood-thinners/

• https://uihc.org/health-topics/complete-blood-count-guide-patients-cancer

• https://www.donatingplasma.org/donation/donor-faq

Bone Marrow Aspiration vs. Bone Marrow Trephine Biopsy

Bone marrow is a smooth, rubbery material that exists within bones and creates new blood cells. Bone marrow creates white blood cells, red blood cells, and platelets, so bone marrow health is essential to fight diseases in the body, as well as function properly. Physicians typically collect bone marrow samples to examine the presence of cancer. Because bone marrow is within the bones, physicians tend to use this biopsy method for cancers that impact the bones such as lymphomas, leukemias, and myeloma. 

There are two different methods for collecting bone marrow samples, including bone marrow aspiration and bone marrow trephine biopsy. Both methods collect bone marrow samples through the back of the hip bone or breast bone and are conducted by an oncologist, hematologist, or another trained technologist. A bone marrow biopsy or aspiration are conducted for numerous reasons, including: 

• Discover the presence of cancer

• Discover the presence of blood disorders such as leukemia, anemias, multiple myeloma, or polycythemia vera

• Find infections or tumors

• Help discover the best course of treatment for the patient

• Stem cell transplantation or chromosomal analysis

Here, we’ll provide an overview of different biopsy methods and information on the overall procedure. Our biorepository provides bone marrow samples from healthy patients that have been collected and store by licensed and certified health professionals. Learn more about our bone marrow samples and order today. 

Bone Marrow Aspiration

Bone marrow aspiration is conducted when a doctor or specialized nurse collects the liquid bone marrow in one piece. The health care professional conducting the test will inject a needle with a tube attached to it, which will then collect a small sample of bone marrow fluids. These fluids are sent to a laboratory and examined under a microscope. The cells are analyzed for blood disorders to discover cancer or infection that has spread to the bone marrow.

Bone Marrow Trephine Biopsy

A bone marrow biopsy differs from aspiration by collecting a solid sample of bone marrow as well as a liquid sample. After the liquid sample is collected (in the same manner it is collected during an aspiration), a hollowed needle is injected. This hollow needle is moved deeper into the bone and collects a tiny sample bone with the marrow inside. 

What to Expect During a Bone Marrow Examination

A bone marrow biopsy or aspiration is conducted by a physician that specializes in blood disorders or cancer. In some instances, a specialized nurse will conduct the test. While the bone marrow exam usually takes about 10 minutes to complete, however, the preparation and post-procedure care can take longer. You can expect to lay on your stomach or side on top of an examination table or hospital bed. If needed, they will provide sedation to help calm you and will numb the site of the test. Your medical professional will give you instructions on caring for yourself after the procedure.

Potential Risks

After your procedure, it’s normal to feel some discomfort or pain from the needle insertion site. Some people may feel pain down the back of their leg, and mild bruising is often seen as well. 

If you notice any of the following symptoms, however, contact your doctor. 

•  Fever of 101 degrees Fahrenheit

• Bleeding that does not easily stop 

• Unusual discharge from the needle insertion site 

• Pain that doesn’t go away at the needle insertion site

Don’t be scared to ask your doctor questions before your procedure. They can provide you the information needed and any specifics about the procedure. 

Resources:

• https://medlineplus.gov/ency/article/003658.htm

• https://medlineplus.gov/ency/article/003934.htm

• https://www.uofmhealth.org/health-library/hw200221

• https://www.uofmhealth.org/health-library/hw200221#hw200228

• https://www.cancerresearchuk.org/about-cancer/cancer-in-general/tests/bone-marrow-test

• https://www.cancer.net/navigating-cancer-care/diagnosing-cancer/tests-and-procedures/bone-marrow-aspiration-and-biopsy
  
  

Frozen Bone Marrow: Cryopreservation

Bone marrow is a spongy tissue and fluid that lives inside your bones, generating blood cells. Bone marrow tests help oncologists and health professionals check if cancer cells are present, and are typically conducted for cancers that are most likely to affect bone marrow such as lymphoma, leukemia, and myeloma. This process can be done as a diagnostic tool or to see how well your cancer treatment is working.

Bone marrow creates the following, all of which are essential for your health: 

• Red blood cells: Red blood cells carry oxygen throughout the body. 

• White blood cells: White blood cells assist in fighting infection and disease threatening the body. 

• Platelets: Platelets help blood clot when a wound is present, helping in controlling the bleeding. 

Cryopreservation Protocol

An autologous bone marrow transplant is when your bone marrow is harvested from your own marrow, which is then frozen for future use. Autologous is self-donated bone marrow, compared to allogeneic transplants that are taken from donors. Our frozen bone marrow samples are collected from certified hospitals and doctors that use healthy donors. We offer a pathology report for every sample that includes age, sex, ethnicity, tumor size, disease stage, procurement date, and treatment history. Order our fresh bone marrow samples today.

The primary applications of frozen bone marrow samples include: 

• Treatment research

• Biomarker tracking 

• Cancer stem cells

• Genetics research

• Single-cell analysis

• Immune response observation

• Toxicology

How is Frozen Bone Marrow Collected

Bone marrow is typically retrieved from your hip bone or the back of your pelvic bone. Frozen bone marrow utilizes a process known as cryopreservation, which has become the standard for biopreservation for its various benefits. Cryopreservation is the process of preserving biological material and storing it below -80°C, which is the temperature of liquid nitrogen. 

During a bone marrow biopsy, a small amount of skin is numbed while a needle is inserted into the bone. The needle collects a sample of blood, bone, and bone marrow, which are studied and examined under a microscope. Once the fresh tissue sample is collected in the operating room, the sample is sent immediately to a pathologist. 

Our geneticists provide a variety of bone marrow samples that are taken from both normal and diseased patients. These samples are taken from the posterior iliac crest with a syringe that contains an anticoagulant and frozen for preservation. Learn more about our frozen bone marrow sampling services.

References:

• https://my.clevelandclinic.org/health/treatments/10866-bone-marrow-harvest-procedure

• https://translational-medicine.biomedcentral.com/articles/10.1186/s12967-019-02136-7

• https://www.cancer.org/treatment/understanding-your-diagnosis/tests/testing-biopsy-and-cytology-specimens-for-cancer/what-happens-to-specimens.html

What is FFPE Tissue Sampling and How is it Used?

Formalin-Fixed Paraffin-Embedded tissue (FFPE) are tissue samples that have been biopsied and prepared for preservation. Formalin is a formaldehyde solution, used for its preservative effects. Paraffin is injected into the tissue sample after the formaldehyde is applied. Paraffin helps to support the tissue sample and protect it from oxidation. FFPE tissue samples help analyze, study, and diagnosing various diseases Additionally, FFPE aids scientists in processing gene expression profiling. 

Standard Protocol for FFPE Tissue 

There is a specific protocol that must be followed to convert regular tissue samples into regular FFPE tissue slides. Here, we will outline the steps to provide you a greater understanding of our process

Fixation 

Fixation is the first step in gathering FFPE tissue slide. Technicians must utilize the correct solution of formalin and paraffin-based on the thickness. The thickness of the tissue sample will determine the quality of the fixed sample. Thicker samples will require more products and more time to reach the center of the tissue.

Dehydration

Paraffin is immiscible with water, so all the water from the formalin solution must be removed from the tissue slide completely. This is done by applying 70% alcohol and slowly increasing to 100% alcohol. Insufficient dehydration of the FFPE tissue sample can lead to tissue degradation, which can harm the process of research and diagnosis. 

Clearing 

This step involves removing the alcohol from the tissue slide with a solution that is miscible with paraffin. Xylene is the most common clearing agent utilized in this step. 

Paraffin Infiltration and Embedding

This step of the process is not standardized, however, our FFPE tissue specimens are collected under IRB approval and conducted by certified medical pathologists. The tissue sample is injected with and surrounded by paraffin. When our technicians conduct FFPE sampling, they fix the tissue in 10% Neutral-Buffered Formalin (NBF) within 30 minutes of surgical excision. Then, the tissue is fixed for 18-24 hours at room temperature, before being embedded in IHC-grade paraffin. Once the paraffin casing has solidified, the FFPE block is ready to be preserved in storage at room temperature. The fixation agent can be customized upon request, and unless indicated otherwise, all FFPE tumor tissue contains at least 50% tumor content. 

Application of FFPE Tissue 

Double-stranded DNA is incredibly stable in FFPE blocks, but less stable molecules such as RNA may degrade within a decade. FFPE is useful because it’s incredibly resilient and can be used for research almost indefinitely, as long as it is stored properly and conducted by a certified and trained professional. FFPE tissue samples include antibodies that bind to specific proteins or structures, allowing doctors to easily identify signs of disease such as cancer or Alzheimer’s.

Alzheimer’s.

• Oncology: FFPE tissue slides are critical to the study of oncology. Preserved tumor tissue showcase characteristic morphologies and specific proteins that are used in diagnosis. 

• Hematology: FFPE tissues is critical when studying blood-related disorders. Hematology has helped scientists and doctors discover cures for numerous diseases caused by abnormalities in the blood. 

• Immunology: Studying tissue samples from a person with an autoimmune disease, such as Lupus or Celiac disease, allows scientists to understand the cause of the disease. Once the cause of the disease is identified, they can create a medication or treatment for that disease. 

• Comparative Research: Tissue from healthy donors is imperative for research. Researchers can compare diseased or unhealthy tissue with healthy tissue, which is critical for the development of new treatments and the future of medicine.   

Explore our FFPE tissue blocks inventory in our biorepository.

What’s the Difference Between FFPE Tissue and Frozen Tissue? 

While we provide frozen tissue services, FFPE has a few advantages that frozen tissue does not provide. However, frozen tissue still provides advantages and benefits in some situations. Speak with a professional to identify which test would be best for you. 

• Storage: FFPE tissue slides can be stored and studied for years, and require more cost-effective methods. Frozen tissue requires expensive freezers that require energy and maintenance. 

• Durability: Due to their fragility, frozen tissues can be difficult to preserve long-term. The tissue deteriorates at a faster rate than FFPE tissue. Frozen tissue degrades quickly while in room temperature environments. 

• Uses: Tissue samples that require staining and morphology study, FFPE tissue samples are most beneficial because they can be handled in various studies. 

A primary disadvantage of frozen tissues is that they are not allowed to thaw after initial freezing, however, they work very well for molecular genetic analysis. Learn more about our frozen tissue sampling services. 

Sources:

• https://lab-ally.com/histopathology-resources/ffpe-samples/

• https://www.biochain.com/general/ffpe-vs-frozen-tissue-samples/

• https://www.biochain.com/general/what-is-ffpe-tissue/