Trans FI/V EDTA Vacuum Blood Collection Tubes C€ IVD Safety Data Sheet can be obtained at www.caltag.co.uk TransFix/EDTA Vacuum Blood Collection Tubes (TVTs) are intended for collection and storage of human whole blood specimens for immunophenotyping of white blood cells by flow cytometry. Recovery of lymphocyte subset markers can be accomplished over a 14-day period following collection. TVTs are for In-Vitro Diagnostic Use. Immunophenotyping by flow cytometry provides a rapid and accurate assessment of the frequency and type of leucocytes in a blood sample. Any delay in testing the sample, such as transport from the collection site to analysis location, can have a negative impact on results. Sample preservation provides a solution which addresses delays between sample collection and testing. TVTs consist of purple capped polyethylene terephthalate tubes that are designed for direct-draw blood collection. TVTs contain a solution of TransFix and K3EDTA at the correct volume to simultaneously stabilise and anti-coagulate human whole blood at the point of collection. Subsets of leucocytes can be distinguished by cell surface antigens using fluorescent antibodies and flow cytometry. Qualitative and quantitative changes in leucocyte subsets are used to identify and monitor immunodeficiency and haematological diseases [1]. The TransFix stabiliser acts by preserving the cell surface antigens of lymphocyte subsets until processing and analysis can be performed. TVTs are available in two sizes: a 3 mL and 9 mL final draw volume tube. The vacuum contained within the TVT ensures that the TransFix reagent is administered at the correct ratio of 1 part TransFix to 5 parts whole blood (1:5). TVTs are sterilised by ionising radiation. 1. TVTs are intended for use as specified in this document. They are in-vitro diagnostic medical devices for professional use only. 2. TVTs are only for use with flow cytometry analysis. 3. Under-filling of tubes will result in an incorrect blood-to-additive ratio and may lead to incorrect analytic results or poor product performance. 4. Do not freeze the TVTs, or blood specimens collected in TVTs. 5. Incubation times or temperatures other than those specified may lead to erroneous results. 6. Do not use TVTs after the expiration date on the tubes and packaging. 7. Only use TVTs to collect human whole blood specimens. Do not use tubes for collection of materials to be injected into patients. 8. Do not dilute or add other components to TVTs. 9. TVTs should only be used by trained phlebotomists. 10. Do not transfer specimens that have been collected in other tubes or specimens treated with other preservatives / anticoagulants into TVTs. 11. Do not use cell viability stains on blood collected in TVTs as they are fixed instantaneously. 13. TransFix treated blood and all materials coming into contact with it should be handled as if capable of transmitting infection. TVTs contain TransFix and the anticoagulant, K3EDTA. TransFix is a clear green liquid containing formaldehyde and other chemicals. H317 - May cause an allergic skin reaction. P261 - Avoid breathing fume, mist, spray, vapours. P280 - Wear protective gloves, protective clothing, eye protection. P302+P352 - IF ON SKIN: Wash with plenty of soap and water. P308+P313 - IF exposed or concerned: Get medical advice/attention. P362+P364 - Take off contaminated clothing and wash it before reuse. Since TVTs contain chemical additives, it is important to avoid possible backflow from the tube. To guard against backflow: 1. Keep patient's arm in the downward position during the collection procedure. 2. Hold the tube with the cap in the uppermost position so that the tube contents do not touch the stopper in the cap or the end of the needle during sample collection. 3. Release tourniquet once blood starts to flow in the tube, or within 2 minutes of application. Indications of Deterioration in unused TVTs 1. Cloudiness or precipitate visible in the TransFix. 2. Colour change of TransFix from a clear green liquid. 3. TransFix change from liquid to solid. If indications of product deterioration occur, do not use TVTs and contact Caltag immediately on: +44(0)1280 827460 or [email protected]. TVTs should be disposed of in accordance with local regulations. Avoid disposing into drainage systems and the environment. Once the TVT contains a biological sample it must be disposed of in accordance with local regulations regarding clinical waste. Storage Conditions and Stability All TransFix products are shipped at ambient temperature (2 - 25°C). Additional insulation may be required for shipment during extreme temperature conditions. TVTs are supplied in a sealed foil pouch. Tubes in an unopened or opened pouch must be stored at 2 - 8°C until the expiration date on the label. Materials needed but not supplied • Phlebotomy materials for sample collection • Automatic pipettes with disposable tips for 10, 20, 100 and 1,000 pL. • Flow cytometry tubes • Flow cytometry calibration and control reagents • Specific fluorescent antibodies and isotype controls • Red blood cell lysing solution • Automatic agitator/vortex machine • Flow cytometer 1. Collect blood by venepuncture. TVTs are compatible with shielded needle devices from most major manufacturers. 2. Fill tube completely. Blood will be aspirated up to the correct total volume and no further. This is important to avoid an incorrect TransFix to blood ratio that could affect results. 3. Remove the TVT from the needle holder and immediately mix by gentle inversion 10 times to distribute the T ransFix throughout the blood sample. Inadequate or delayed mixing may result in coagulation and inaccurate test results. Do not vortex. 4. After collection, store/transport the blood filled TVT at 2 - 8°C for up to 14 days or for up to 3 days at 18 - 25°C. 5. If refrigerated, incubate the TVT at room temperature (18 - 25°C) for 15 minutes prior to use. 6. Mix the TransFix treated blood by rolling the TVT between the hands 10 times and by inverting 10 times. Heavier cells and blood components will sediment over the 14 day period, forming two distinct layers; this is normal. 7. Remove and reinsert the cap by grasping with a simultaneous twisting and pulling action, not by a 'thumb roll' method. 8. Perform immunophenotyping by flow cytometry in accordance with the manufacturer's instructions. A 'stain, lyse-no wash' sample preparation method is recommended. 9. TVTs containing blood should be returned to storage at 2-8° C within 6 hours for future use, if necessary. 10. When treated with Transfix the dilution factor must be accounted for when calculating absolute cell counts. Adjust the absolute cell count by multiplying the output by 1.2. 1. It is recommended that all antibody conjugates are validated in association with TransFix prior to use. 2. Use caution when implementing automatic gating strategies as light scatter positions of cells stabilised by TransFix may differ from those of untreated cells. 3. Gating for lymphocytes should be performed on CD45+/SSc.