This Application Note describes an antibody-based method for identifying and counting blood cells from whole blood samples on the Agilent 2100 bioanalyzer without a washing step. The ability of the Agilent 2100 bioanalyzer to identify peripheral whole blood cells is demonstrated and compared to a flow-cytometry-based reference instrument.
This document details an antibody-based method for identifying and counting peripheral whole blood cells on the Agilent 2100 bioanalyzer. The study demonstrates that the device can distinguish red blood cells (RBCs), total white blood cells (WBCs), and specific WBC subpopulations like granulocytes and monocytes without requiring a washing step or scattering data.
The note provides specific staining protocols for RBCs using anti-human CD235a, WBCs using SYTO 16 nuclear acid dye with a lysis reagent, and WBC subpopulations using dual-fluorescent markers (anti-CD14 and anti-CD16). Results show that the Agilent 2100 bioanalyzer can successfully analyze these populations and provide cell counts comparable to established automatic blood cell counters.
Yes, the document describes an antibody-based method to identify and count blood cells from whole blood samples on the Agilent 2100 bioanalyzer without a washing step.
The monoclonal antibody anti-human CD235a conjugated with the fluorochrome Alexa 647 is chosen for RBC staining.
WBC subpopulations are studied using two types of fluorescent markers: a blue dye (SYTO 16) and red fluorescent monoclonal antibodies (anti-CD14 and anti-CD16 conjugated with PE-Cy5).