Absolute counts of peripheral blood leukocytes were determined for all study participants using BD Trucount tubes, six-color TBNK Reagent and, in addition, CD123 BUV395 (BD Biosciences), CD15 PB, CD193 BV605, and HLA-DR BV785 (Biolegend), and CD14 PE-Cy5 (eBioscience) according to the manufacturer’s instructions. Samples were fixed in 1× BD fluorescence-activated cell sorter (FACS) lysing solution (BD Biosciences) for 2 h and acquired on a BD FACSymphony A5 instrument, equipped with ultraviolet (355nm), violet (405 nm), blue (488 nm), yellow/green (561 nm), and red (637 nm) lasers. For absolute cell count calculations, the number of events for populations of interest was divided by the number of bead events and multiplied by the BD Trucount bead count.
Fluorescence activated cell sorter facs lysing solution
Manufactured by BD
The BD fluorescence-activated cell sorter (FACS) lysing solution is a laboratory reagent designed to prepare cells for analysis or sorting using flow cytometry. The solution is used to lyse red blood cells while preserving the desired cell population, allowing for more accurate analysis and sorting of the target cells.
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Lab products found in correlation
Cd123 buv395, by BD (1 mentions)
Cd14 pe cy5, by Thermo Fisher Scientific (1 mentions)
Cd15 pb, by BioLegend (1 mentions)
Cd193 bv605, by BioLegend (1 mentions)
Facsymphony a5 instrument, by BD (1 mentions)
Hla dr bv785, by BioLegend (1 mentions)
Trucount tube, by BD (1 mentions)
Quantibrite pe beads, by BD (1 mentions)
Percp cy5, by BioLegend (1 mentions)
Phycoerythrin pe, by BD (1 mentions)
Facscalibur cytometer, by BD (1 mentions)
Fluorescein isothiocyanate (fitc), by BioLegend (1 mentions)
2 protocols using fluorescence activated cell sorter facs lysing solution
1
Multicolor Flow Cytometry Leukocyte Profiling
2
Quantifying CD64 Expression on Leukocytes
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Blood samples were collected using ethylenediaminetetraacetic acid-2Na tubes and kept at room temperature. Within 24 hours, 50 μL of whole blood aliquots were stained with anti-human antibodies against the pan-leukocyte marker CD45 (clone H130) conjugated with PerCP-Cy5.5 (BioLegend, San Diego, CA, USA), anti-human CD16 antibody (clone 3G8) conjugated with FITC (BioLegend), and anti-human CD64 antibody (clone 10.1) conjugated with phycoerythrin (PE) (BD Pharmingen). After the samples were incubated for 20 minutes in the dark at room temperature, we added 1 mL of BD Fluorescence-activated Cell Sorter (FACS) Lysing Solution (BD Biosciences). The samples were then incubated for another 20 min or stored at 4°C overnight. Neutrophil and monocyte populations were gated by side scatter and CD45 fluorescence intensity, and the geomean of CD64 was then assessed by flow cytometry using a FACSCalibur cytometer (BD Biosciences). We constructed a standard curve using QuantiBRITE® PE beads (BD Biosciences) to calculate the number of CD64 molecules per cell.
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