1. Serum separation tubes (SSTs) are tubes used in phlebotomy containing a silicone gel; when centrifuged the silicone gel forms a layer on top of the buffy coat, allowing the blood serum to be removed more effectively for testing and related purposes. 0000020480 00000 n Do not remove the tube stopper.
Do not disturb the cell layer or carry any cells over into the pipette. For routine collections, use a 21- to 22-gauge needle. (See Figure 10.) 0000010908 00000 n (See Figure 13.
0000024913 00000 n With some needle assemblies, you may slide the collection tube into the holder, carefully pushing the tubes forward until the needle touches the stopper. The essential difference between plasma and serum is that plasma retains fibrinogen (the clotting component), which is removed from serum. <>>> %%EOF
Learn how and when to remove this template message, "Experimental setup for the ultrasonic fractionation of flowing whole blood in a capillary", Statement by Dr. Kathryn Zoon, Food and Drug Administration, https://en.wikipedia.org/w/index.php?title=Blood_fractionation&oldid=1136318874, Short description is different from Wikidata, Articles needing additional references from March 2013, All articles needing additional references, Creative Commons Attribution-ShareAlike License 3.0, This page was last edited on 29 January 2023, at 19:57. 0000001651 00000 n A needle length of 1 to 1 inches permits an angle of entry that will not pierce both vein walls and enter tissue. 1 0 obj (See Figure 11.). If multiple samples are being drawn, invert each specimen as soon as it is drawn. SST tube 30 minutes. 10. If specimen is centrifuged before clotting is complete, a fibrin clot will form on top of the cell. Allow the puncture site to air dry after swabbing, or dry the site with gauze. Serum Separator Tubes (Gold Top) Serum separator tubes contain a clot activator and a 0000005797 00000 n Allow specimens collected in a clot tube (eg, red-top or gel-barrier tube) to clot before centrifugation. =MLJ#9 Spreader slide pushed across the slide in a jerky manner. If you are using your own collection equipment instead of the vacuum tube technique, use only clean, dry, sterile needles, syringes, and tubes. WebThe color of the lowest layer of centrifuged blood may appear dark red or bright red depending on the oxygen content of the cells. 2 0 obj After collection of the whole blood, allow the blood to clot by leaving it undisturbed at room temperature. Address this problem by collecting a fresh tube when blood flow is established or select another puncture site and, using sterile/unused equipment, collect a second specimen. Do not apply fixative. 0000002286 00000 n First, it is helpful to establish some baseline definitions: Blood samples intended for laboratory analysis are usually collected in glass or plastic tubes that have been partially evacuated so that their internal air pressure is lower than atmospheric pressure. Complete blood count samples are drawn in a purple- or lavender-topped EDTA tube. Careful attention to routine procedures can eliminate most of the potential problems related to specimen collection. The purple/lavender top Vacutainer tube contains EDTA, an anticoagulant. This usually takes 1530 minutes. The most common serum preparation considerations: Separate serum from red cells within two hours of venipuncture. This usually takes 1530 minutes. This can lead to changes in concentration of certain analytes. WebSerum separation tubes (SSTs) are tubes used in phlebotomy containing a silicone gel; when centrifuged the silicone gel forms a layer on top of the buffy coat, allowing the blood serum to be removed more effectively for testing and related purposes. 0000025995 00000 n
No portion of the film should extend to the edges of the slide. 
This process is sometimes called cold alcohol fractionation or ethanol fractionation. 12. Anticoagulants and Preservatives. 
If the glass syringe has been autoclaved, it should be oven dried before use. Fasting or diet restrictions, such as low-fat diets, should be explained in detail, particularly to aged or overanxious patients or their caregivers.
Do not remove the needle from the vein with the vacuum tube engaged.
When drawing blood, please follow all venipuncture procedures recommended for use by recognized organizations and/or in accordance with applicable state regulations involving phlebotomy practices.
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Select two clean, grease-free glass collection slides with frosted ends (new ones whenever possible). For additional information regarding preparation of serum samples, view the following video: 1. For most profile testing, draw at least two 8.5-mL gel-barrier tubes. Common causes of a poor blood film. Troponin I is a positively charged molecule, and there has been speculation that the highly negatively charged polymeric heparin effectively shields Troponin I from being recognized by antibodies. Use: Serum or clotted whole blood. Also, use of a blood pressure cuff, in lieu of a tourniquet, will reduce trauma to fragile red blood cells. Be sure to provide the laboratory with the amount of serum specified. The kit has everything you need for fingerstick blood diagnostic tests and blood sample collection devices. 
(See Figure 7.). [2][3] Human serum albumin prepared by this process is used in some vaccines, for treating burn victims, and other medical applications. Remove and invert each successive tube after it is filled. Pipette the serum or plasma into a clean plastic screw-cap vial Unless otherwise indicated, serum specimens may be sent at room temperature.
Normal serum (far left) followed by icteric specimens ranging from 1+ to 4+, In all specimens, the normal serum is shown on the left, followed by the abnormal serum specimens; 1) Jaundice/Icterus, 2) Lipemia, 3) Hemolysis; http://clinical-laboratory.blogspot.com/2013/06/preventing-pre-analytical-errors.html. Either plasma or serum may be separated from the blood cells by centrifugation. 
When specimens are not collected in the basal state, the following additional effects should be considered when interpreting test results. After drawing blood, observe proper venipuncture techniques to prevent continued bleeding and/or hematoma. Plasma Preparation Using a Plasma Preparation Tube (PPT). The test request form and all specimens are sent in one container (box or plastic specimen transport bag). If the sample type you wish to test is not listed in the IFU, consult the manufactures technical support staff as it is likely someone else has asked the same question before you. 4. If the patient cannot be taken off the medication in question, its presence should be noted on the test request form. Gently tap tubes containing additives to dislodge any material that may be adhering to the stopper. Do. 0000017471 00000 n
Leave it in that position. WebIf it turned red colour, we could be explain the hemolysis will occur when animal test. 
Do not reuse needles. Blood glucose determinations may be ordered at a specific time to check the effect of insulin treatment. Web3) Serum separator tubes (Tiger top) can be substituted for red top tubes in some instances but should be avoided for certain endocrinology and clinical pathology tests such as a progesterone baseline, thyroid panels, cortisol tests and certain drug levels. 86 0 obj
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There are four steps involved in obtaining a good quality specimen for testing: (1) preparation of the patient, (2) collection of the specimen, (3) processing the specimen, and (4) storing and/or transporting the specimen. The resulting supernatant is designated serum. NOTE: All drug levels must be drawn in red top tubes only. Prepare the second film in the same manner. Assembling Supplies. WebA machine called a centrifuge spins your blood to separate your red blood cells, platelets and plasma. Thiazide diuretic therapy. WebSerum Preparation. %PDF-1.6
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Failure to keep or have the spreader slide at approximately a 45 angle. 0000017685 00000 n
For example, RBCs have 20x more potassium (K) than serum/plasma, so prolonged contact and/or contamination with red cells will significantly change K levels. Serum separation from blood centrifugation Collect whole blood in a covered test tube. Prolonged centrifugation may cause hemolysis. 0000032207 00000 n
Oral contraceptives cause a decrease in serum vitamin B12 levels that is often indistinguishable from vitamin B12 deficiency of any cause. Avoid hemolysis: red blood cells broken down and components spilled into serum. 0000004728 00000 n
Webred serum after centrifugation. Send plasma in a plastic transport tube labeled Plasma, EDTA. Send whole blood in a lavender-top tube. Provide sufficient quantity of specimen to meet the minimum fill line on preservative transport container. An overnight fast is preferable (from 6 PM of the evening previous to collection) to ensure that the patient is in the basal state. Most cases of hemolysis can be avoided by observing the steps listed. The dextrose is a nutrient for some of the cells in the blood, and the citric acid complexes with or chelates calcium which prevents the blood from clotting, Minutes after collection, the blood is centrifuged into packed RBCs and citrated plasma for better storage. 4. WebSpecimens collected in tubes that do not contain a gel separator must be separated after centrifugation by physically removing the supernatant plasma or serum with a pipet and transferring to a plastic aliquot tube. Before you collect. Recommended times are based upon an intact clotting process. The laboratory sends out a report marked QNS (quantity not sufficient), and the patient has to be called back for a repeat collection at an inconvenience to the patient and to the physician.
Remove the tube carefully without disturbing the contents. As the blood is separated, the heavier reds cells sink to the bottom and are given back to you. Please follow the instructions that come with the kit and call your LabCorp representative if you have any questions. Plasma is the fluid component of blood. WebA machine called a centrifuge spins your blood to separate your red blood cells, platelets and plasma. Invitrogen has not evaluated some of these tubes for compatibility with our ELISA kits. NOTE: All drug levels must be drawn in red top tubes only. This can be a valuable resource in the process of developing an assay.
Mix by inverting specimen with additive immediately after collection. This can cause discomfort to the patient and may also cause hemolysis. Several different types of anticoagulants interfere with the activity of these factors to prevent clotting. After allowing clot to form, insert the tube in the centrifuge, stopper end up. Ordinarily, a 21- or 22-gauge needle is used. They also cause an increase in total serum thyroxine-binding globulin. Slide preparation using venous blood is described below. If blood is drawn for routine hematology, prepare the blood films (blood smears) immediately after collection.
Put the Frozen Specimen Keeper containing the specimens in your lockbox according to the pictorial instructions provided (see link above). 11. Before centrifugation, the sample must be completely solidified.
Centrifuge the specimen at the end of the waiting period in accordance with the manufacturer's instructions for speed. SST II Vacutainer with clot activator gel AFTER centrifugation, separating the blood cells (bottom) from the serum (top). Regardless of diet and length of fast, some patients may produce cloudy specimens. Recommended times are based upon an intact clotting process. Hold the left corners of the collection slide with the left thumb and forefinger.
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13. Store unpreserved specimens refrigerated or in a cool place until ready for transport. Following the collection, preparation, and transport instructions suggested by LabCorp supports the best possible test results. 7. 0000017494 00000 n
Use: Sodium fluoride whole blood or plasma. WebRed cells (RBCs) often have a much higher concentration of analytes than the liquid portion (serum/plasma) of blood. Complete the test request form to indicate time and date of collection along with collector's identification. These containers can keep frozen specimens frozen, but they will not be able to freeze specimens at room temperature or refrigerated specimens. Use blood transfer device to fill tubes from syringe. If there is difficulty in performing venipuncture, minimum volume may be submitted if it is indicated in the test description. 0000045947 00000 n
Click hereto learn more. If commercially available tubes are to be used, the researcher should use the red topped tubes. The gel-barrier tube is an additive tube and should be inverted five to six times after collection. When all samples have been drawn, remove the entire assembly from the arm. Frozen Serum. Why do we need to separate the serum from the red blood cells after centrifugation? Please refer to the specimen requirements for the test(s) of interest available in the Directory of Services. Allow specimens collected in a clot tube (eg, red-top or gel-barrier tube) to clot before centrifugation. 
WebThe effect of recentrifugation of serum separator tubes on concentration of serum analytes In clinical laboratories, sometimes there is a need to recentrifuge the original tubes ("clot" tubes) in order to better clarify and clean the serum or plasma for further analysis. 0000016260 00000 n
After mixing, store the BD PPT upright at room temperature until centrifugation. The most common urine collection considerations: Collection of Vacuum Tubes Containing Additives (eg, anticoagulants, preservatives, clot activators, gel-barrier). Before centrifugation, the sample must be completely solidified. Many common prescription and nonprescription (over-the-counter) medications can interfere with chemical determinations or alter levels of substances measured. It is important to gain the patient's understanding and cooperation in obtaining an acceptable specimen. Maintain and transport the specimen at the temperature indicated in the test requirements. The red cells are delicate, round, concave bodies that contain hemoglobin, the complex chemical that transports oxygen and carbon dioxide. When using vacuum tubes containing an additive: Collection of Vacuum Tubes Without Anticoagulants.
Turn the centrifuge off, if not an automatic turn off, and allow it to come to a complete stop. Prolonged centrifugation may cause hemolysis. Only one test request form accompanies the serial samples, and it is completed with all patient information, including any medications administered and the number of samples sent. To ensure an adequate specimen volume: The definition of specimen temperatures for storage and shipping is as listed below: Frozen: -1.0 to -80.0 oCFrozen Specimen Guide. Pipette the serum or plasma into a clean plastic screw-cap vial WebThe color of the lowest layer of centrifuged blood may appear dark red or bright red depending on the oxygen content of the cells. Collect in an appropriate collection tube. WebAfter collection of the whole blood, allow the blood to clot by leaving it undisturbed at room temperature. The clot is removed by centrifugation and the resulting supernatant, designated serum, is carefully removed using a Pasteur pipette. NOTE: All drug levels must be drawn in red top tubes only. All Rights Reserved. The practices below help ensure acceptable samples for all analytes. The cells are removed by centrifugation. Well-prepared films can be used in performing the differential white cell count, for examining the morphology (size, structure, and shape) of red and white cells to determine the presence of abnormal cells, and also for the examination of the size and number of platelets. G7P3a['vu2zKgDy8HL&UhAqiHxAb7T 04 &-s'/Ev+;d"iU*SYYG@9W;psED0jRr Mt}=5FWfw`r:hJ&( (dKCNJ>FI|Ii Small amounts of moisture can cause hemolysis. It is quick and easy to get excellent separation of centrifuged blood with the aid of a high-quality blood separation centrifuge such as the CAPPRondo Advanced Clinical Centrifuge CRC-416X. Avoid hemolysis or red blood cell breakdown. 5gGdaQgJs=fDvU%.ao4gdkjN2g@>pn5j,6L]-; 8W yFHEri$8wt`tj# 5S_PFsZN"MqZg,:X0Z?> {Ef:zl3B~v*^c+EKtfbmj5 R/5vgOc{ ]Jh-9~. Anchor the vein. To avoid dietary-induced high lipid levels prior to testing, many physicians require patients to exclude the high-fat foods from their diets or to fast for 12 to 14 hours prior to specimen collection. 3 0 obj It is important to separate the cellular and liquid portions of a blood specimen as soon as possible when the test requires a sample of serum or plasma.
Inform patients that fasting does not include abstaining from water. Whole blood, serum, and various plasmas are not interchangeable sample matrices. If any preservative powder is visible, continue inverting the tube slowly until the powder is dissolved. 
Web3) Serum separator tubes (Tiger top) can be substituted for red top tubes in some instances but should be avoided for certain endocrinology and clinical pathology tests such as a progesterone baseline, thyroid panels, cortisol tests and certain drug levels. 7. Single Specimens. The composition of blood is altered after meals by nutrients being absorbed into the bloodstream. It is used for many tests in the clinical laboratory. Let the specimen stand for 30 to 60 minutes and (preferably) not longer than 60 minutes prior to centrifugation. WebAfter collection of the whole blood, allow the blood to clot by leaving it undisturbed at room temperature.
This requirement is important in order to achieve the proper blood-to-additive ratio; otherwise, the specimen may be found to be QNS. WebSerum separation tubes (SSTs) are tubes used in phlebotomy containing a silicone gel; when centrifuged the silicone gel forms a layer on top of the buffy coat, allowing the blood serum to be removed more effectively for testing and related purposes. The blood should not extend past 3/4 the length of the slide. HWne*:jIH G@I(7{i{OU5# MSf#l^Zvl?l/ _|qo_,|u3O~]^m)Uw+ad9=mKJu_J9NaJxu>{zxv&o_p?U. To avoid this assay interference, EDTA plasma and the alkaline phosphatase containing label must never come in contact for any length of time. Dispose of the used needle and holder in a sharps container according to the provisions in your exposure control plan. (See. endobj It is important to avoid freeze-thaw cycles because this is detrimental to many serum components. When using vacuum tubes containing no additives: In general, grossly or even moderately hemolyzed blood specimens may not be acceptable for testing. Add the preservative (as specified in the test requirements) to the urine collection container prior to collection of the specimen if the preservative is not already in the container. WebThe effect of recentrifugation of serum separator tubes on concentration of serum analytes In clinical laboratories, sometimes there is a need to recentrifuge the original tubes ("clot" tubes) in order to better clarify and clean the serum or plasma for further analysis. Treat all biological material as material that is potentially hazardous as well as contaminated specimen collection supplies. Hemolysis occurs when the red cells rupture and hemoglobin and other intracellular components spill into the serum.
As soon as possible after collection, set the blood upright in a test tube rack. Send whole blood in a yellow-top tube. Serum is usually collected in mottled red/gray, gold, or cherry red-top tubes, and red-top tubes are occasionally used.
For all those who are involved in specimen collection and preparation, the responsibility to adhere to current recommendations designed to maintain the safety of both patients and health care workers does not end when the patient is dismissed. Do not disturb the cell layer or carry any cells over into the pipette. (See Figure 5.) Greasy or dirty slides, or use of a spreader slide with a chipped or unpolished end. Heparinized tubes (green tops) are indicated for some applications; however, heparin can often be contaminated with endotoxin, which can stimulate white blood cells to release cytokines. Plain tubes with no anticoagulants have red stoppers and are used in the preparation of serum after clotting and centrifugation. Do not remove the stopper. WebRed cells (RBCs) often have a much higher concentration of analytes than the liquid portion (serum/plasma) of blood. As an alternative to energy-consuming centrifugation, more energy-efficient technologies have been studied, such as ultrasonic fractionation.[1]. Fasting here is defined as no caloric intake for at least eight hours. Plain tubes with no anticoagulants have red stoppers and are used in the preparation of serum after clotting and centrifugation. Quattrocchi F,Karnes HT,Robinson JD,Hendeles L.,Ther Drug Monit. a) Mature erythrocytes (red blood cells), b) polymorphonuclear segmented neutrophil (white blood cell), c) eosinophil (white blood cell), d) basophil (white blood cell); Also seen on the slides are platelets. There are numerous coagulation factors (factor VIII, factor IX, etc) involved in the clotting of blood. Keep the spreader slide at a 45 angle, maintaining light but firm pressure with the spreader slide against the horizontal slide. Gel-barrier Tubes. Remove the clot by centrifuging at 1,0002,000 x g for 10 minutes in a refrigerated centrifuge. The aseptic method of collecting and transporting a blood specimen works on the principle of a vacuum tube for drawing blood. Whole blood contains the liquid fraction of blood (i.e., plasma) as well as the cellular elements that lead to clotting under certain circumstances. Refer to individual test descriptions for specific information. When blood is collected, it fills a plastic bag that contains a solution of dextrose and citric acid. WebSerum separation tubes (SSTs) are tubes used in phlebotomy containing a silicone gel; when centrifuged the silicone gel forms a layer on top of the buffy coat, allowing the blood serum to be removed more effectively for testing and related purposes. To test the effect of a certain medication, a physician may order the same test to be obtained on consecutive days, before, during, and after the patient has received a medication. When multiple tests requiring frozen serum are ordered, a plastic transport tube should be prepared for each test. Purple or lavender top tubes contain EDTA, and green top tubes have heparin in them. (See Figure 14.). Provide patients with adequate containers and instructions for 24-hour urine and stool collections. The samples should be maintained at 28C while handling. 0000014326 00000 n 2. These tubes have color-coded polymer stoppers that indicate their contents. If you are a user of an assay, you should consult the manufacturers instructions for use (IFU) to determine what sample types have been validated for use in the assays. Too much pressure in drawing blood into a syringe or forcefully ejecting blood into a collection tube from a syringe may damage red cells. 144 0 obj <>stream Provide the proper mixing of specimen with urine preservative as specified in the test requirements. Most laboratory testing for clinical purposes is done on samples obtained from blood. 0000030089 00000 n 113 0 obj <> endobj This step embeds the full point of the needle in the stopper without puncturing it, preventing blood leakage on venipuncture and the premature loss of vacuum. Remove the tube carefully without disturbing the contents. When frozen serum is required, transfer the serum using a pipette into a plastic transport tube. Slides must not be touched on any area except the long slide edges or frosted ends.
Mix by inverting specimen with additive immediately after collection. Note: If a patient has intravenous (IV) solutions going into one or both arms, it is acceptable to puncture the vein 3 to 4 inches below the site of the IV. Platelets are small fragments of special cells that aid in blood clotting. Plasma proteins are separated by using the inherent differences of each protein.
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