Information

Blood and plucked-feather sample storage


This next field season I will be collecting both blood and feather samples and I wondered how best to store the samples. The blood samples will be used for microsatellite and/or SNP analysis. The feather samples will be used for genetic sexing and/or microsatellite analysis.

Feathers: can these be stored in envelopes at "room" temperature? I've seen mixed answers in the literature. They are plucked feathers and so may have a bit of blood or tissue at the tip. If they are used for microsatellite analysis, I want to ensure this DNA is not degraded. I will be storing them frozen at my field site, but our partners at other sites may not have access to freezers.

Blood: we will be collecting blood into capillary tubes. I have a few Whatman FTA cards, but not enough. So I'm planning to freeze the blood in the capillary tubes. Are there better options than this? I'm planning to use non-heparinized tubes. Out of curiosity, has anyone used blood from FTA cards for microsatellite or SNP analysis? Can I expect sufficient DNA quantity and quality? Bird DNA is nucleated, so I should get more bang for my buck (volume-wise).

I appreciate your help!


Collection and Storage DNA Evidence

Each year, Genetic Technologies, Inc. successfully analyzes several cases from evidence vaults dating back to the early 1970’s. Investigators and others gathering DNA evidence should do so with great care, utilizing the following guidelines.

DNA, in a dry state, is very stable. Ultra-violet light, extreme heat and high humidity are the primary destructive agents of the DNA molecule. There are a few basic rules that must be followed for the proper collection, packaging and storage of DNA evidence.


Current best practices of cold storage of biological samples

In the United States alone, there are more than 40,000 individual research laboratories located on university campuses that are advancing the field of biological and biomedical sciences. Researchers within these laboratories have assembled a very large collection of biological samples from clinical and field studies, some irreplaceable, all representing enormous scientific and financial value for the researcher and the organization (universities, research institutes, biotechnology/pharmaceutical companies, biobanks, etc.). The cost per sample collected can range from a few dollars up to $10,000. 1 There are currently over a billion samples (DNA, RNA, cells, clones, tissue organs, blood, buccal swabs, etc.) collected and warehoused in thousands of research labs and bio-banks globally. These samples are of high value to researchers, and current research trends are driving growth of these collections at an escalating rate.

To preserve these important research assets, organizations and individual researchers engaged in biological and biomedical research invest a huge sum of money in capital equipment purchases and maintenance of cold storage facilities to stabilize and store their large inventory of samples. However, there are increasing disadvantages to this method. For example, cold refrigeration/freezers produce hydrofluorocarbons, which are some of the most potent greenhouse gas pollutants 2 with a deleterious impact on the environment. Quantitatively, according to a report in The Economist, the typical ultra-low-temperature freezer consumes about 7,665 kWh per year while releasing 54,805 pounds of carbon dioxide. This is equal to the emission from about four cars. 3

Additional challenges to the use of cold storage are highlighted below:

  • Cold packing and shipping produce a large amount of waste materials.
  • Purchase costs, maintenance costs and energy costs of cold refrigerators/freezers add up as sample collection grows, and accelerate as the cost of energy increases.
  • Heat generated from refrigerators/freezers further adds demands to facility requirements, costs and planning to stabilize environmental conditions at lower temperatures than would be required without the equipment.
  • Freezers take up an increasingly large amount of lab space, potentially inhibiting current and new facility/ research space needs.
  • Multiple freeze-thaw cycles can lead to sample quality degradation.
  • Cold freezing, especially of bio-tissues, can lead to cell membrane damage.3
  • Power failure or freezer failure can place samples at risk for degradation and loss.

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2 METHODS

2.1 Collection of DBS samples in the field

SHARE collected DBS samples during its Wave 6 in 2015 in 12 European countries and Israel. We used harmonized collection protocols and DBS collection kits, thoroughly trained our interviewers and implemented interviewer monitoring throughout the fieldwork. All DBS samples arriving at the central biobank in Denmark for storage were visually inspected for number of blood spots, spot quality (smeared or overlapping, as these change the distribution of analytes in the filter paper) and spot size (small spots indicate that the interviewer did not wait until a sufficiently large blood drop had formed before collecting the blood on the filter paper). Further shortcomings noted included missing desiccant (influencing humidity protection) and spot discoloration (indicating that a wet DBS was packaged). Other uncontrollable impacts on the samples were malfunctioning national postal systems, with consequences on shipment time, and unusually high temperatures encountered during shipment. The implementation and monitoring of the DBS collection in SHARE Wave 6 has been described (Börsch-Supan et al., 2020 ).

2.2 Validation structure

The aim of the validations is to simulate the environmental conditions and fieldwork effects experienced by DBS and to then measure the resulting change in biomarker values in a structured approach. The environmental data collected during fieldwork monitoring and by sample inspection enabled us to design the validation scheme.

Figure 1 shows a conceptual model of the structural differences between VBS collected in a laboratory, DBS collected in the field and its mirror image, our VBS-based DBS validation scheme structured by the same processes.

Standard VBS analyses are depicted in the center of Figure 1. For a given marker, the laboratory analysis yields a value Y that corresponds to the reference value based on serum or whole blood, depending on analyte.

The DBS field process is depicted in the left side of Figure 1, marked in blue. A donor provides capillary blood from a finger-prick, which is dropped onto a filter paper card and creates spots of varying size (dependent on the volume of the blood drop DBS_C). This blood is dried and then experiences the various conditions found to arise during fieldwork and shipment such as differences in temperature, humidity, and drying and/or shipment time (DBS_F). In the lab, the dried blood is liquefied and assayed, yielding a value of V for the marker.

Our validations mimicked the field process but used venous whole blood to create DBS in order to obtain sufficient material for all 171 treatments from a single donor, which would not be feasible using finger pricking to obtain capillary blood. This process is depicted on the right side of Figure 1, marked in red. The donor's venous blood was collected in an EDTA-coated tube, pipetted onto a filter paper card and dried (DBS_V), and then treated under replicable laboratory conditions to simulate the typical fieldwork and shipping treatments described above (ie, heat, humidity, etc. DBS_T). The dried blood was then liquefied and analyzed to yield the value X. For the validation experiment we also collected capillary DBS from each donor (DBS_C), yielding the value W, as well as analyzed untreated DBS_V, resulting in the value Z.

The outcomes X, Y, Z, and W are different due to three groups of effects generated by chemical and physical processes. These effects are shown in green in Figure 1.

First, the blood source is either capillary blood from a finger-prick or venous blood collected by cannula. Capillary blood is a heterogeneous mixture of plasma, interstitial fluid and blood cells while venous blood does not contain interstitial fluid. Centrifugation of venous blood to obtain serum removes all cellular material. In addition, venous blood analyzed directly or used to obtain plasma for analyses, is treated to prevent coagulation, for example, with EDTA. There are also small differences between plasma and serum in the concentrations of some analytes. We call the implications of these differences between capillary and venous blood on the assays the “sample-type effects.” These effects have not been extensively studied for the analytes of interest here. Spooner, Ramakrishnan, Barfield, Dewit, and Miller ( 2010 ) reported different marker values for blood collected from finger prick, venous cannula and whole blood cannula when measuring paracetamol exposure. They speculated that the observed lack of interchangeability between the sampling sites may not be limited to drugs.

Second, the capillary blood is dried on filter paper. During the drying process, cell lysis occurs and releases cellular content. The dried blood is re-liquefied in the lab for analysis. These processes may influence the chemical composition of the blood and the marker molecules. We call this the “dry-liquefy effects”.

The combination of sample-type and dry-liquefy effects creates differences between the outcome values W, Y, and Z. The difference between Y and Z has been extensively studied by Crimmins et al. ( 2014 ) and is 3-fold for most analytes: the difference between serum and whole venous blood, the treatment of the latter with EDTA, and it subsequent drying and reliquefaction. Isolation of the dry-liquefy effect would involve comparisons between dried/eluted plasma spots and whole blood that had undergone sonication, or freeze/thaw, to release cellular contents such comparisons are beyond the aims of this study. The difference between Y and W is a combination of the blood source (capillary whole blood vs venous serum) and the dry-liquefy effect. The difference between W and Z has two causes: the difference between capillary blood and venous blood, and the EDTA-treatment of the latter.

The third group of effects are the original and simulated “fieldwork effects” (heat, humidity, etc.), depicted as blue and red rhombi in Figure 1. These effects have not been systematically investigated previously and are the focus of this study.

Our aim is to establish a conversion formula following this structure that is applicable to the SHARE population and estimates the value that we would have obtained had it been feasible to analyze donors' venous blood by standard analytical methods for serum, plasma or whole blood.

More formally, the aim of this study is to establish an equation, Y = f(X, T, H), which computes the estimated standard value for Y from the treated DBS value X, the applicable treatment conditions T (eg, temperature, humidity, drying time, shipment time, spot size) and donor characteristics H (eg, health, age, sex) reflecting potential interaction effects (Section 3.3). Ideally, this conversion formula would have accuracy comparable to the measurement variation of the reference value Y obtained from serum or venous whole blood (Woodworth et al., 2014 ).

2.3 Data

The validation study and all marker assays were performed at the University of Washington Department of Laboratory Medicine and Pathology Biomarker Laboratory (UW).

Over a period of 10 days (July 20 to 30, 2018) venous blood was collected under laboratory conditions by a phlebotomist from 20 donors recruited by BloodWorks NorthWest, Seattle. WA. Venous blood was collected into K2EDTA vacutainers and into tubes without added preservatives or stabilizers. Immediately after collecting the venous blood, the DBS were created. Whole blood was stored at ambient temperature and was couriered to the UW laboratory within 4 hours of collection. Once received, EDTA plasma and serum by clot was separated by centrifugation at 3000 rpm for 10 minutes. Liquid whole blood (for HbA1c and tHb), serum and plasma samples (for all other markers) were created and assayed the same day. Analyses of the serum created the venous reference marker values, called Y in Figure 1. Venous EDTA blood samples were further processed as follows:

(i) We created DBS samples from EDTA whole blood (DBS_V). They were immediately frozen at −70°C after drying and kept frozen until analysis, resulting in the value Z

(ii) Of the blood from the 20 donors, we created DBS samples that were subsequently exposed to 171 different controlled conditions (DBS_T in Figure 1) mimicking the fieldwork conditions that we encountered during the SHARE Wave 6 DBS collection, resulting in the value X.

At the time of the venous blood collection, the phlebotomist also collected a capillary blood sample from a finger-prick for DBS using the same technique applied during SHARE fieldwork (DBS_C). After drying and transfer to UW the DBS cards were frozen at −70°C. Filter-paper cards used to create the DBS were the same as those used during SHARE fieldwork: Ahlstrom 226 filter paper. Ahlstrom 226 is fully comparable to the Whatman 903 protein saver cards predominantly used in the US and by HRS. The CDC found no difference between the performance properties of 903 vs 226 filter papers, which also produced comparable results across analytes and testing methods (Mei et al., 2010 ). Desiccant used during SHARE and in the simulations of shipment times and storage was 2 g molecular sieve in a Tyvek pouch (Absorpower Service GmbH, Germany). Analyses of the DBS_C resulted in the value W.

  • Spot size: We created dried blood spots in three sizes, using 10 μL, 30 μL or 60 μL of blood, respectively, as a large percentage of the field-collected spots had diameters < 1 cm (60 μL).
  • Drying time: The samples were dried at room temperature for 5, 20, 35, or 240 minutes and then immediately packed as for shipment. The short drying times reflect frequent survey circumstances which did not permit overnight drying the long drying time refers to full dry (Mei, 2014 ).
  • Humidity protection:
  1. Closed polyethylene (PE) bag: The samples were put into PE bags, which then were zip-closed or left open.
  2. Desiccant: A desiccant was either placed inside the PE bag or not.
  • Outside temperature: The samples (inside their PE bags and envelopes) were exposed to 5°, 20°, or 35°C for 2 hours. This mimics the exposure to (high) outside temperature under the assumptions that the fieldwork samples were exposed to the actual outside temperature for only a fraction of the shipment time.
  • Shipment time: The samples were left at room temperature for additional 3, 7, or 14 days.

Our data set contains 3420 observations clustered by 20 donors, each including a certain treatment combination, the respective values for the analyzed biomarkers (see below) and three values from different kinds of “untreated” blood samples: venous blood (VBS), DBS samples from venous EDTA-whole blood (DBS_V), and DBS samples from capillary blood (DBS_C). “Untreated” in this sense means that they were of optimal size (60 μL) and were immediately frozen after the optimal drying time (240 minutes), hence, they were created under laboratory conditions as opposed to the DBS that were subjected to the simulated varying fieldwork conditions described above. The capillary DBS samples will be part of the conversion equation used for recalculating the fieldwork samples, which are actually capillary blood DBS, not venous blood DBS as in this validation study. In addition to the blood values, we had information on age, gender and body mass index (BMI) score for each donor.

The liquid-blood assays for the VBS were performed on a Beckman-Coulter Olympus AU680 Chemistry Analyzer for HDL, TC, TG, and CRP CysC was analyzed with a microtiter plate assay. THb was assayed using a Sysmex XN hematology analyzer. All HbA1c tests, both on whole blood and DBS samples, were run on a Bio-Rad Variant II High Pressure Liquid Chromatography (HPLC) System.

For the analyses of the DBS samples, UW has developed standard (in-house) assays based on the above-mentioned references. For those assays, validations have shown roughly a coefficient of variation of 5% to 10% in inter-assay, inter- and intra-spot reproducibility. To keep the inherent variation of our validation samples low, for each assay all samples from a single donor were assayed together.

2.4 Statistical analysis

We also estimated a simplified model as well as a larger model which includes all treatments and all possible interactions (up to 6-fold). The six estimated models are summarized in Table 1.

Model name basic simple allint basicH simpleH allintH
Treatment variables x x x x x x
Single interactions x x
Squared treatment variables x x
All possible interactions x x
Gender (dummy) x x x
BMI score x x x
BMI score squared x x x
Age (years) x x x
Age squared x x x


Discussion

In our analysis, we observed that the ethical issues arising from the collection, export and reuse of samples are inter-related. Thus, there is the need to discuss and understand these issues from the perspectives of all research actors and in the context of the research interactions between host research institutions and local communities (what we refer to as micro-level issues) and interactions between collaborating institutions (macro-level issues). Also, we did not find any major differences between the different sexes and age groups in our analysis of the data. The major differences in opinion had more to do with the number of years respondents had been exposed to research and their roles in the research institution.

Our data suggest that biomedical research is evolving rapidly and its success in the future will increasingly depend on access to human biological samples and collaborative partnerships. This is consistent with current literature on biobank research highlighting the important societal value of biobanks in advancing biomedical research [8, 27–30]. It is also suggested that biomedical research is a complex and expensive enterprise requiring the pulling together of expertise and resources to achieve important research goals which include alleviating suffering, advancing knowledge, preserving life and promoting human well-being. Thus, research collaborations will continue to be a key way of moving both the science and ethics of research forward [5, 31, 32]. Our research supports the findings of previous empirical studies in Africa in identifying a high level of general support for biomedical research activities in Africa [17, 33, 34]. However, there is a pressing need for a number of practical ethical concerns to be addressed in order to ensure high standards of practice and maintain public confidence in international research collaborations, particularly those involving the collection, export and reuse of human biological samples.

At the micro-level are local concerns about the use of blood samples in research which have resulted in rumours about too much blood being taken, blood-selling and devil worshipping, as reported in the past in these research settings [19, 35]. Interviewees in this study attributed these concerns in part to the perceived lack of understanding, unfamiliarity, uncertainties and complexities associated with novel research projects such as genetic and genomic research. Thus one proposed solution is to identify innovative and effective ways of communicating the rationale for these scientific research practices to all research actors including fieldworkers, community representatives and research ethics committees to allay these local fears. However, it was also evident that there are deep cultural sensitivities around the use of blood in general which need to be taken seriously. Although we did not seek to compare our findings with non-African perspectives, our data suggests that cultural sensitivities around blood samples are more pronounced in these African settings because of their historical background and cultural attachment to blood. The concerns raised in this study support other claims in the literature linking historical accounts with people’s perception of medical research in general and the local community’s view of some research institutions in Africa as ‘blood-stealing’ organizations [35–37]. As these scholars have also suggested, these cultural sensitivities reveal how local communities are responding to their relationships of dependence and inequality with host research institutions [37]. This raises issues of justice and benefit-sharing as ethical concerns, which require serious attention in determining whether research is ethical or not.

To address concerns about the validity of broad consent, many stakeholders interviewed have proposed the need to identify innovative ways of meaningfully engaging sample donors and their communities, particularly on decisions around the future uses of samples. Clearly, with the growing recognition that research with human biological samples have implications for the wider community and population, most of these suggestions are calls for extending the ethical principle of respect for persons to communities as well. It is worth noting that both KWTRP and NHRC have established mechanisms for communicating with local residents about the institutions’ research activities. The processes involved in these CE strategies have been reported elsewhere [18, 38, 39], and include regular interactions with networks of voluntary community representatives to consult with, public meetings with the general public and community leaders in local villages, community member visits to the research institution, and fieldworker support and training [39, 40]. Interviewees in both sites were confident that these engagement activities are effective ways of improving research literacy and strengthening the trust relationship between the host institutions and the community. In Kilifi for example there are on-going in-depth consultative activities with a diverse range of community members on appropriate forms of benefit-sharing for research, including for studies involving blood sampling [41, 42]. There are also examples of deliberative processes that have proved useful in soliciting public views that could be explored in specific contexts [43, 44]. However, what remains unclear is the extent to which sample donors and communities should be involved in decisions around future uses of samples. Further empirical studies on what methods of community engagement will be most effective in this process would be desirable.

At the macro-level, this research highlights concerns among African researchers and research ethics committees on the fate of exported samples and who stands to gain in research collaborations. These responses also suggest that the issue about local control of research samples and proper recognition does not disappear because the collaboration is just between African institutions. What is important is the nature of the relationship, especially one of trust, that is developed between the various research actors in the collaboration. While some stakeholders have suggested the need for strengthening the capacity of host research institutions to enable much of the research process to be conducted locally and to enhance local control of samples, it was also evident that no level of local capacity can completely eliminate sample export. These include requirements for uniformity of analysis, quality control and the growing demands for sample and data sharing. Researchers’ concerns are also based on principles of justice and calls for some assurance that, irrespective of where research takes place, the interests of the less dominant partners in the collaboration (host African institutions, participants and communities) will be adequately protected, against the background of inevitable sample export. This can also be achieved through clear, transparent and fair research agreements [45, 46] as well as ensuring feedback and accountability on the fate of exported samples. These processes are important to bolster host communities’ confidence in the research enterprise and to protect participants and communities from the effects of misplaced trust.


Collecting and Storing Tissue, Blood, and Bone Marrow Samples From Patients With Rhabdomyosarcoma or Other Soft Tissue Sarcoma


Condition or disease Intervention/treatment
Adult Rhabdomyosarcoma Childhood Desmoplastic Small Round Cell Tumor Chordoma Desmoid-Type Fibromatosis Metastatic Childhood Soft Tissue Sarcoma Non-Metastatic Childhood Soft Tissue Sarcoma Previously Treated Childhood Rhabdomyosarcoma Recurrent Adult Soft Tissue Sarcoma Recurrent Childhood Rhabdomyosarcoma Recurrent Childhood Soft Tissue Sarcoma Rhabdomyosarcoma Stage I Adult Soft Tissue Sarcoma AJCC v7 Stage II Adult Soft Tissue Sarcoma AJCC v7 Stage III Adult Soft Tissue Sarcoma AJCC v7 Stage IV Adult Soft Tissue Sarcoma AJCC v7 Untreated Childhood Rhabdomyosarcoma Other: Cytology Specimen Collection Procedure Other: Laboratory Biomarker Analysis

I. Collect human tumor tissue and other biological specimens (blood, serum, and bone marrow) from patients with rhabdomyosarcoma or non-rhabdomyosarcoma soft tissue sarcoma diagnosed and/or treated at a Children's Oncology Group (COG) member institution.

II. Provide a repository for storage of tissue and other biological specimens collected by COG investigators from these patients.

III. Make these specimens available for approved projects by laboratory-based investigators.

IV. Collect clinical data on these patients who are not being treated on a COG therapeutic study.

V. Define and compare the clinical features of patient subgroups with alveolar rhabdomyosarcoma whose tumors carry the t(213), t(113) or neither translocation.

VI. Investigate the relationship between evidence of submicroscopic disease and response rate (CR/PR), failure-free survival, and survival of patients with alveolar rhabdomyosarcoma, as determined by positive or negative reverse transcriptase-polymerase chain reaction (RT-PCR) assay for the t(2:13) and t(1:13) on peripheral blood and bone marrow specimens obtained at diagnosis.

VII. Compare the clinical, cytogenetic, and molecular biologic features of patient subgroups with anaplastic rhabdomyosarcoma and other subtypes of rhabdomyosarcoma.

Surgical tissue, bone marrow, and blood specimens are collected at diagnosis (initial or relapse) and, if applicable, at the development of a second primary tumor. Specimens are used for research purposes. A certificate of confidentiality protecting the identity of research participants in this project has been issued by the National Cancer Institute.

Patients who are not enrolled on a Children's Oncology Group treatment trial are followed every 6 months for at least 10 years or until disease progression or development of a second malignancy.

Layout table for study information
Study Type : Observational
Estimated Enrollment : 150 participants
Official Title: A COG Soft Tissue Sarcoma Diagnosis, Biology and Banking Protocol
Actual Study Start Date : March 15, 1999

Resource links provided by the National Library of Medicine
  1. Collection of human tumor tissue and other biological specimens (blood, serum, and bone marrow) from patients with rhabdomyosarcoma or non-rhabdomyosarcoma soft tissue sarcoma [ Time Frame: Up to 10 years ]
  2. Collection of clinical data on patients who are not being treated on a COG therapeutic study [ Time Frame: Up to 10 years ]
Information from the National Library of Medicine

Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the contacts provided below. For general information, Learn About Clinical Studies.

Layout table for eligibility information
Ages Eligible for Study: up to 50 Years (Child, Adult)
Sexes Eligible for Study: All
Accepts Healthy Volunteers: No
Sampling Method: Non-Probability Sample

Histologically or cytologically confirmed diagnosis of 1 of the following:

Non-rhabdomyosarcoma soft tissue sarcoma

  • Chordoma
  • Desmoid fibromatosis
  • Desmoplastic round cell tumors
  • Undifferentiated embryonal sarcoma of the liver
Information from the National Library of Medicine

To learn more about this study, you or your doctor may contact the study research staff using the contact information provided by the sponsor.

Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT00919269

Show 247 study locations Hide 247 study locations
  • Specimen collection requires withdrawing blood, cerebrospinal fluid, collecting urine, or swabs from mucosal surfaces.
  • Specimen collection is performed using aseptic techniques to ensure sterility of the sample and avoid contamination from bacteria or other bodily fluids.
  • The types of biological samples accepted in most clinical laboratories are: serum samples, virology swab samples, biopsy and necropsy tissue, cerebrospinal fluid, whole blood for PCR, and urine samples. These are collected in specific containers for successful processing in the laboratory.
  • PCR: polymerase chain reaction
  • necropsy: The pathological dissection of a corpse particularly to determine cause of death. Applicable to the examination of any life form.
  • biopsy: The removal and examination of a sample of tissue from a living body for diagnostic purposes.

Laboratory diagnosis of an infectious disease begins with the collection of a clinical specimen for examination or processing in the laboratory.

The laboratory, with the help of well-chosen techniques and methods for rapid isolation and identification, confirms the diagnosis.

It has been observed that the most important and frequent factor affecting laboratory analysis, even in a well-functioning laboratory, is not the laboratory investigation itself but specimen preparation and errors in identification or labeling. Proper collection of an appropriate clinical specimen is, hence, the first step in obtaining an accurate laboratory diagnosis of an infectious disease.

Applying one&rsquos knowledge of microbiology and immunology for the collection, transportation and storage of specimens is as important as it is in the laboratory. For starters, the interpretation of the observation may be misleading if the specimen is inadequate.

There are several types of specimens recommended for diagnosis of immunological diseases including: serum samples, virology swab samples, biopsy and necropsy tissue, cerebrospinal fluid, whole blood for PCR, and urine samples.

Serum is the preferred specimen source for serologic testing. Blood specimens are obtained aseptically using approved venipuncture techniques by qualified personnel. Specimens are allowed to clot at room temperature and then are centrifuged. Serum is transferred to tightly-closing plastic tubes and stored at 2 &ndash 8°C before shipment&ndashwhich should always be prompt. Acute serum should be collected at the onset of symptoms. Convalescent specimens should follow two to four weeks later. Paired sera are tested together.

Figure: Venipuncture: Performed to draw blood sample using a vacutainer

Plasma is also collected for a very limited number of tests. Lipemic, hemolyzed, or contaminated sera may cause erroneous results and should be avoided as should repeated freeze-thaw cycles.

Another type of specimen used for disease diagnosis is cerebrospinal fluid (CSF). This should be transported in tightly-closing plastic tubes. Refrigerated CSF is acceptable for a limited number of serologic tests however, if PCR is to be performed for the viral panels, the specimen must be frozen and shipped on dry ice. CSF specimens should be clear of any visible contamination or blood. A lumbar puncture (or LP, and colloquially known as a spinal tap) is performed to collecte CSF. This consists of the insertion of a hollow needle beneath the arachnoid membrane of the spinal cord in the lumbar region to withdraw cerebrospinal fluid for diagnostic purposes or to administer medication.


The Musculoskeletal System

The musculoskeletal system provides support to the body and gives humans (and many animal species) the ability to move. The body&rsquos bones (the skeletal system), muscles (muscular system), cartilage, tendons, ligaments, joints, and other connective tissue that supports and binds tissues and organs together comprise the musculoskeletal system.

Most importantly, the system provides form, support, stability, and movement to the body. For example, the bones of the skeletal system protect the body&rsquos internal organs and support the weight of the body. The skeletal portion of the system serves as the main storage depot for calcium and phosphorus. It also contains critical components of the hematopoietic system (blood cell production). The muscles of the muscular system keep bones in place they also play a role in movement of the bones by contracting and pulling on the bones, allowing for movements as diverse as standing, walking, running, and grasping items. To allow motion, different bones are connected by joints. Within these joints, bones are connected to other bones and muscle fibers via connective tissue such as tendons and ligaments. Cartilage prevents the bone ends from rubbing directly on each other. Muscles contract (bunch up) to move the bone attached at the joint.

Figure (PageIndex<1>): Joints, tendons, and ligaments: To allow motion, different bones are connected by joints. Within these joints, bones are connected to other bones and muscle fibers via connective tissue such as tendons and ligaments. Figure (PageIndex<1>): Human muscular system: The muscles of the muscular system keep bones in place while assisting with movement by contracting and pulling on the bones.

Unfortunately, diseases and disorders that may adversely affect the function and overall effectiveness of the system exist and can be detrimental to the body. These potentially debilitating diseases can be difficult to diagnose due to the close relation of the musculoskeletal system to other internal systems. In humans, the most common musculoskeletal diseases worldwide are caused by malnutrition. Ailments that affect the joints, such as arthritis, are also widespread. These can make movement difficult in advanced cases, they completely impair mobility. In severe cases in which the joint has suffered extensive damage, joint replacement surgery may be needed.

Figure (PageIndex<1>): Human skeletal system: The bones of the skeletal system protect the body&rsquos internal organs, support the weight of the body, and serve as the main storage system for calcium and phosphorus.

Progress in the science of prosthesis design has resulted in the development of artificial joints, with joint replacement surgery in the hips and knees being the most common. Replacement joints for shoulders, elbows, and fingers are also available. Even with this progress, there is still room for improvement in the design of prostheses. The state-of-the-art prostheses have limited durability, wearing out quickly, particularly in young or active individuals. Current research is focused on the use of new materials, such as carbon fiber, that may make prostheses more durable.

Figure (PageIndex<1>): Prostheses: Improvements in the design of prostheses, artificial replacements for body parts such as joints, elbows, legs, and fingers, have allowed for a wider range of activities in impaired recipients.


Storing blood from mouse - Can blood be stored, and if yes, How? (Aug/26/2005 )

I've got a question regarding storing blood obtained from mouse so that the red blood cells can be used the next day for binding studies. Currently, I'm unable to get viable cells for the binding assay if I store the blood overnight. Can someone please give me some good advice on how to go about handling and storing murine blood for use on another day? Sincere thanks!

Many eons ago I used to perform CBC/chemistry at a diagnostic lab. We stored all of our blood at 4 degrees for up to one week in the event we needed to rerun something.

Whole blood is best stored with clotting inhibitor (ie. EDTA) in tube at 4 degrees. Don't freeze whole blood. Breakdown of RBCs will occur over time, so it is best to use ASAP. Serum or plasma can be frozen at -80 degrees until needed, but should be isolated immediately to prevent hemolysis.

On that note, hemolysis can also occur while obtaining the sample and handling it. I would consider spinning down one tube as a test to see if the hemolysis is due to sample prep or to the O/N storage.

Many eons ago I used to perform CBC/chemistry at a diagnostic lab. We stored all of our blood at 4 degrees for up to one week in the event we needed to rerun something.

Whole blood is best stored with clotting inhibitor (ie. EDTA) in tube at 4 degrees. Don't freeze whole blood. Breakdown of RBCs will occur over time, so it is best to use ASAP. Serum or plasma can be frozen at -80 degrees until needed, but should be isolated immediately to prevent hemolysis.

On that note, hemolysis can also occur while obtaining the sample and handling it. I would consider spinning down one tube as a test to see if the hemolysis is due to sample prep or to the O/N storage.

Is the above also valid for blood from mouse? human blood can be stored for extended periods if stored appropriately . what about mouse blood? has anyone been using stored RBCs from mouse blood after storage overnight? Thanks.