I am counting cells from haemolymph by flow cytometry . i want to know how these true count beads will be helpful in calculation and analysis
Dear Preethy,
The BD Truecount tubes contain fixed and known number of beads to which you need to add your sample to it. Your cells can be counted by comparing the cell events to the bead events either by software or manually. For more details follow the instructions from the kit.
http://www.bdbiosciences.com/ds/is/tds/23-3483.pdf
As the name suggests, these tubes will give you the TruCount of the cell types your looking at since they have a fixed numbers of beads. Generally, approximately 50 uL of blood is added by reverse pipetting (to avoid the pipetting error) followed by the recommended volume of the specific antibody and vortexed. If your not worried about the RBC count, u can simply add lysing solution after an antibody binding incubation period of 20 minutes. 15-20 minutes after adding the lysing solution, u can acquire the cells in a flow cytometer. For an automated cell count, use a clinical software like the FACS CANTO S/W.
Thanks for all your kind response . I need actual calculation using true count beads
Hi Preethy,
I haven't used the "TruCount", but I have used the "Count123" beads from Ebio. Are you asking how to calculate? Here is how I do it. See attached.
Nate
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