Blood Collection Procedures

Choose method and schedule for blood collection. Common survival blood collection methods for metabolic profiling are tail-nick, tail snip, saphenous vein, submandibular (cheek), and retroorbital bleeding. With the tail nick or tail snip blood collection methods, 75 pi samples of blood can be collected up to four times in a 1-day experiment, not exceeding a total of 250 pi, and are used as default blood collection procedures. This usually provides up to 30 pi serum samples and is sufficient to measure glucose, insulin, and other metabolic markers. In some cases, larger... 

At 7.5, 15, 30, and 60 min after glucose injection, repeat the blood collection procedure. 

Assess blood collection procedure (cytokine release by leukocytes and cytokine stability in biological fluids)... 

Several references for these variables—species, gender, and age—are also listed in Appendix A. Referenced compendiums of data often show diverse values for a single laboratory species obtained by different investigators and techniques, and this diversity is in part due to the use of different blood collection procedures and analytical techniques (Mitruka and Rawnsley 1977 Caisey and King 1980 Matsuzawa, Nomura,... 

Blood collection procedures can have a significant impact on study results, and a variety of collection sites is used, particularly for the smaller laboratory animals. Blood sampling procedures require skilled personnel, and it is not uncommon to see a higher incidence of poor-quality specimens when routine procedures have not been used or the operator is not sufficiently proficient. Some variations can be traced to individuals performing the collection procedures in slightly different ways... 

Several of the blood collection procedures alter the plasma albumin and total protein concentrations, and these changes should be considered when interpreting toxi-cokinetic data, particularly when perturbations of plasma proteins occur (Hulse et al. 1981 Chou and Levy 1984 Tamura et al. 1990 Proost, Wierda, and Meijer 1996). 

The blood collection procedure that has been in use in the authors laboratory involves drawing blood with a stainless steel needle into a plain glass vacuum tube (No. 6430, Becton-Dickinson and Co., Rutherford. NJ 07070). The blood is allowed to clot approximately 20 min before centrifugation. Serum is transferred to a 17 x 100 mm polypropene tube (Falcon. Oxnard, CA). In establishing a reference range for healthy individuals, blood specimens from 50 subjects were collected by this method with a resulting aluminium range of 1 to 12 [Pg.285]

Stainless steel needles and plastic catheters used in blood collection contribute negligible amounts of aluminum to the procedure. Standard collection equipments, e.g., Vacutainer and Monovetten , can be used after checking all sources for possible contamination. Separation aids must be avoided. The blood collection procedure in the authors laboratories involves drawing blood with a stainless steel needle into an EDTA-K Monovette (Sarstedt, Numbrecht, ERG). After centrifugation the plasma samples are directly transferred to the vials of the atomic absorption spectrometry (AAS) sampler. 

Kluft C., Verheijen J. H. Leiden Fibrinolysis Working Party. Blood collection and handling procedures for assessment of tissue-type plasminogen activator (t-PA) and plasminogen activator inhibitor-1 (PAI-1). Fibrinolysis 1990 4(Suppl2) 155-61. 

Blood collection from the tail vein is a simple and rapid, nonsurgical method which does not require anesthesia. A relatively large number of serial samples can be obtained within a short period of time. However, this method is limited to relatively small sample volumes (<250 pi per sample). Although larger volumes can be obtained by placing the rat in a wanning chamber, this procedure could significantly influence the disposition of the test compound and therefore is not recommended for routine studies. Blood collected from the cut tail has been shown to provide valid concentration data for numerous compounds. 

Different procedures are described for pre-treatment of blood, but perchloric acid is the most widely used agent blood collected in a tube containing heparin is immediately deproteinised with a perchloric acid solution (1 mol/1) that has been refrigerated at + 4°C (two volumes for one volume of blood). Deproteinised samples may be frozen for up to 5 days. 

New Zealand rabbits are normally used for serum production they are easily handled and adapt well to individual cages or group floor pens. This strain has half-lop ears, which make blood collection from marginal veins a fairly straightforward procedure. 

All animal experimental procedures (including handling, housing, husbandry, blood collection, and drug treatment) must be conducted in accordance with national and institutional guidelines for the care and use of laboratory animals. 

Measure baseline glucose by tail tip or tail snip blood collection, according to procedure described above. 

Before beginning the terminal collection procedures have all serum separation tubes for blood collection labeled and all tubes for tissue collection labeled and filled with 5-10 ml 10% formalin (3.7% formaldehyde). Organs from the same mouse can be pooled in the same tube as they can simply be embedded in a single block for histological analysis. 

Table 9 summarizes procedures of blood collection for aluminum determination. Steps to avoid contamination, the analytical technique, and the aluminum levels found in the serum of healthy individuals are listed. It can be seen that, as a general procedure, plasticware was used, all materials that entered in contact with the sample were acid-washed, usually by nitric acid, and the analyses were carried out inside clean rooms. 

The site for blood collection has to be different from the site of intravenous administration. Thus, the vena cephalica antebrachii can be used for collection and the vena saphena parva for administration (or vice versa). The dog is kept by one person and the hairs at the selected venes at the forelimbs or hindlimbs are removed by gently clipping. The vein on the dog s limb is compressed by the assistant by encircling the limb with the hand or applying a rubber tourniquet loop. The puncture procedure is then performed. Blood should be withdrawn slowly by aspirating gently. 

Samples Obtained from Biological Fluids As illustrated in Figure 5.1, biological fluids, which also have two compartments, are placed in group I. Such fluids include blood (often classified as a tissue), urine, semen, tears, and saliva. Many of these fluids contain cells as a normal component, while in others the cells represent a contamination. Fluids like saliva that contact the outside and, when collected, often contain microbes, are examples of contaminated materials. Sometimes such microbes are the result of the collection procedure, and their numbers can often be controlled by careful technique. At other times, as is the case with urine, their presence can indicate an underlying disease process. In any case, the study of enzymes from such fluids again requires the separation of the two compartments. 

Gross, M.D. Prouty, C.B. Jacobs, D.R.J. 1995. Stability of carotenoids and a-to-copherol during blood collection and processing procedures. Clin. Chem. 41 943-944. 

In a few patients, backflow from blood tubes into veins occurs owing to a decrease in venous pressure. The dangerous consequences of this occurrence may be prevented if only sterile tubes are used for collection of blood. Backflow is minimized if the ai m is held downward and blood is kept from contact with the stopper during the collection procedure. To minimize problems if backflow should occur and to optimize the quality of specimens- especiaUy to prevent cross contamination with anticoagulants—blood should be collected into tubes in the following order (1) blood cul-... 

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