A sheath-less combined optical and impedance micro-cytometer

Abstract

We describe a sheath-less micro-cytometer that measures four different parameters, namely fluorescence, large angle side scatter and dual frequency electrical impedance (electrical volume and opacity). The cytometer was benchmarked using both size and fluorescent bead standards and demonstrates excellent size accuracy (CVs ≤ 2.1%), sensitivity and dynamic range (3.5 orders of magnitude) at sample flow rates of 80 μL per minute. The cytometer was evaluated by analysing human blood, and a four part differential leukocyte assay for accurate CD4+ T-cell enumeration was demonstrated. The integration of impedance, fluorescence and side scatter into a single miniature cytometer platform provides the core information content of a classical cytometer in a highly compact, simple, portable and low cost format.