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BD Leucocount™ setup acquisition
BD Leucocount™ setup acquisition

This document describes how to adjust your instrument settings after running BD FACSComp™ software, to acquire your stained BD Leucocount samples.

The method is based on locating the BD Trucount™ beads in the appropriate channels by adjusting the PMT values for FL1, FL2, and side scatter. Also, all compensations are set to zero.

BD Leucocount sample staining and acquisition
BD Leucocount sample staining and acquisition

This is an example of the BD Leucocount report. The sample is stained with the reagent and acquired using BD CellQuest™ Pro software after setting up the system so that the beads are entirely within region R1.

The acquisition is set to stop when 10,000 beads have been acquired. Then the software automatically calculates the final cell concentration based on the correct BD Trucount assayed count from the foil pouch (50,000 in this case).

Generally the regions will not have to be moved to identify the white cells or beads if the setup is done correctly. If there are no residual white cells, there will be no events in R2, so it is important to run controls to verify where to expect them if the blood filter has failed to remove all the white cells.

BD Procount laboratory report
BD Procount™ laboratory report
Stem cell enumeration is important in the transfusion process, in which the physician needs to know the number of stem cells available in the sample to ensure adequate volume for engraftment. The BD Procount reagent stains CD45 and CD34 cells for stem cell enumeration. A nucleic acid dye is added to assist in identifying the nucleated events and for setting the threshold. A control reagent with IgG1 (instead of CD34) is also used with this test, and results are printed on a second page. The IVD software has been validated for use with peripheral blood and fresh apheresis products up to 6 hours after draw. The fixed stained sample can be acquired as soon as 30 minutes from starting staining, or acquisition can be delayed up to 24 hours. The software automatically acquires enough events, performs gating, and calculates and reports. The CD34 cell count, the nucleated (WBC) cell count, and the percentage of white blood cells that are CD34 cells are reported.
Enumeration of CD34+ haematopoietic stem cells
Enumeration of CD34+ haematopoietic stem cells
The BD™ Cell Stem Enumeration kit works with the BD FACSCanto™ system to provide reproducible CD34+ enumeration in a single tube, single platform assay.