Leukocyte differential count is one of the most frequently ordered clinical tests in hospitals. This paper reports a point-of-care test for the leukocyte count by using a microflow cytometer and a fluorescent dye assay. The dye assay relied on fluorescent detection alone to count leukocytes in blood and to identify leukocyte subtypes. By combining the fluorescent assay with a sheathless microflow design, the proposed method achieved a minimal sample volume by eliminating excessive dilution and sheath flow. In this paper, a four-part leukocyte differential count including lymphocyte, monocyte, neutrophil and eosinophil was demonstrated, and the whole test consumed only a small amount of blood (5 μL) and reagents (68 μL in total). The merits of minimal sample volume, long reagent shelf life and portable instrument made this method optimal for point-of-care applications.

This work demonstrated leukocyte count and two-part leukocyte differential from the whole blood based on a portable microflow cytometer system. Leukocytes are selectively stained with a fluorescent dye, Acridine Orange (AO). The blood sample is then pumped through a disposable microfluidic device for fluorescence sensing. Under blue LED excitation (460nm), the green fluorescence from DNA and the red fluorescence from RNA are detected simultaneously with two photomultiplier tubes (PMT). Leukocytes are counted and differentiated into two parts, lymphocyte versus non-lymphocyte, based on their fluorescence signatures. The results, including leukocyte absolute count and lymphocyte ratio, matched well with the commercial hemacytometer counts (maximal error 9.0%, correlation coefficient ~0.8). We also tested the system for the first time under a zero-gravity (zero-G) environment that facilitated its application in space missions.

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This paper reports the four-part differential leukocyte count (DLC) of human blood using a MEMS microflow (flow) cytometer. It is achieved with a two-color laser-induced fluorescence (LIF) detection scheme. Four types of leukocytes including neutrophils, eosinophils, lymphocytes and monocytes are identified in blood samples, which are stained by fluorescein isothiocyanate (FITC) and propidium iodide (PI). The DLC results show good correlation with the count from a commercial hematology analyzer. The whole system is also implemented into a portable instrument for space application.

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