Improved Enumeration of Weakly Fluorescent CD4+ T-lymphocytes by Confining Cells in a Spinning Sample Cartridge with a Helical Minichannel

Abstract

The CD4 (cluster of differentiation 4) counting method is used to measure the number of CD4+ T-lymphocytes per microliter of blood and to evaluate the timing of the initiation of antiretroviral therapy as well as the effectiveness of treatment in patients with human immunodeficiency virus. We developed a three-dimensional helical minichannel-based sample cartridge in which a thread-like microgroove formed in the cylindrical surface and configured a particle-positioning and imaging system equipped with a single DC (direct current) motor that can be controlled by a smartphone application. Confinement and enrichment of CD4 cells within a sharp focal depth along the helical minichannel is accomplished by spinning the cylindrical sample cartridge at high speed before acquiring cell images and thus CD4+ cells with weak fluorescence intensity can be detected even in a channel much deeper than existing two-dimensional flat chambers without an autofocusing module. By detecting more cells in a larger sample volume, the accuracy of the CD4 cell count is improved by a factor of 5.8 with a channel of 500 μm depth and the precision is enhanced by a factor of 1.5 with a coefficient of variation of 2.6%.