Transcription of BD Trucount Tubes
1 1. INTENDED USE. BD Trucount Tubes are used for BD Trucount Tubes determining absolute counts of leucocytes in blood. For determining absolute counts of leucocytes in blood BD Trucount Tubes are designed for use with in vitro diagnostic products such as 25 Tubes Catalog No. 340334 BD Tritest reagents, and a suitably equipped flow cytometer. BD Trucount Tubes can be used with the BD FACS . Loader. 2. PRINCIPLES OF THE PROCEDURE. Procedures described in this instructions for use (IFU) apply to immunophenotyping applications.
2 For 11/2016 23-3483-08 other applications, refer to the appropriate product-specific IFU. IVD. Add the appropriate monoclonal antibody 2016 BD. BD, the BD Logo and all other trademarks reagent and whole blood directly to the are property of Becton, Dickinson and Company. BD Trucount tube. The lyophilized pellet in the tube dissolves, releasing a known number of fluorescent beads. During Becton, Dickinson and Company analysis, the absolute number (cells/ L) of BD Biosciences positive cells in the sample can be 2350 Qume Drive, San Jose, CA 95131 USA determined by comparing cellular events Benex Limited to bead events.
3 If the appropriate Pottery Road, Dun Laoghaire, software, such as BD Multiset , is used, Co. Dublin, Ireland absolute counts will be determined by the Tel + Fax + software. If you are manually performing BD Biosciences data analysis using software such as European Customer Support BD CellQuest Pro , simply divide the Tel + number of positive cellular events by the Fax + number of bead events, then multiply by the BD Trucount bead concentration. Becton Dickinson Pty Ltd, 4 Research Park Drive, Macquarie University Research Park, 3.
4 REAGENT. North Ryde NSW 2113, Australia Becton Dickinson Limited, Each pouch contains 25 BD Trucount 8 Pacific Rise, Mt. Wellington, Tubes , sufficient for 25 tests. Auckland, New Zealand Precautions For In Vitro Diagnostic Use. 1. BD Trucount Tubes are designed for use turned from blue to lavender, discard with a specific lyse/no-wash procedure. the remaining Tubes . Use Tubes within For absolute counting, prepare and 1 hour after removal from the foil analyze samples within BD Trucount pouch and do not use beyond the Tubes .
5 Do not transfer beads to another expiration date indicated on the tube. The presence of proteins, such as packaging. serum proteins contained in whole WARNING All biological specimens and blood, is necessary for proper materials coming in contact with them are performance of BD Trucount beads in considered biohazards. Handle as if absolute Follow specific capable of transmitting infection2,3 and assay IFUs when diluting samples. Do dispose of with proper precautions in not attempt to threshold on forward accordance with federal, state, and local scatter (FSC) for data collection.
6 Do not regulations. Never pipette by mouth. remove the metal retainer in the Wear suitable protective clothing, BD Trucount tube. eyewear, and gloves. It is the responsibility of the user to BD FACS lysing solution is required validate any other method or use. and contains diethylene glycol and The addition of a precise volume of formaldehyde. Refer to the BD FACS. blood is critical to achieving the result. Lysing Solution instructions for use for Pipettes must be calibrated to deliver warnings. exactly 50 L of sample.
7 If this or a similar type of pipette is not used, 4. INSTRUMENT. perform the reverse pipetting technique BD Trucount applications are designed for (see Reverse Pipetting in Section 6 for a flow cytometers equipped with brief description). Refer to the pipette appropriate computer hardware and manufacturer's instructions for more software. The flow cytometer must be information. equipped to detect three-color Always be sure to use the bead count fluorescence, forward scatter (FSC), and from the current lot of BD Trucount side scatter (SSC).
8 We recommend the Tubes when entering this value in the BD FACSC alibur flow cytometer;. software or when manually calculating however, results can be achieved using an absolute count. The correct bead other platforms. Refer to the appropriate count is critical for determining a cell reagent IFU for specific instrument count. Do not mix multiple lots of Tubes limitations. The BD FACS Loader can also in the same assay. be used with this product. BD has developed BD Multiset software, for use Store BD Trucount Tubes in their with specific reagents and BD Trucount original foil pouch at 2 C 25 C.
9 To Tubes , which automatically calculates avoid potential condensation, open the absolute counts. However, you can also pouch only after it has reached room use software such as BD CellQuest Pro for temperature and carefully reseal the data acquisition and analysis and pouch immediately after removing a manually calculate absolute counts. tube. Examine the desiccant each time you open the pouch. If the desiccant has 2. 5. SPECIMEN COLLECTION AND Staining the Cells PREPARATION Stain whole blood samples following Collect blood aseptically by specific instructions in the appropriate venipuncture4,5 into a sterile EDTA reagent IFU.
10 Lyse red blood cells after (lavender top) BD Vacutainer blood staining using diluted (1X) BD FACS. collection tube. Follow the collection tube lysing solution. Use care to protect the manufacturer's guidelines for the Tubes from direct light. Perform the minimum volume of blood to be collected. procedure at room temperature (20 C . Store anticoagulated blood at room 25 C). temperature (20 C 25 C) until ready for Reverse Pipetting staining. A precise volume of whole blood is critical. If a pipette that delivers a precise 6.
