Diffractive Beam Shaper for Multiwavelength Lasers for Flow Cytometry.

Abstract

Illumination spot in a flow cytometer is a crucial factor determining the measurement accuracy and stability. The traditional mechanism is to precisely calibrate multiple optical components to convert circular Gaussian beams into elliptical Gaussian beams, making it difficult to shape multiwavelength lasers simultaneously. A diffractive beam shaper for multicolor lasers with high simplicity, only containing one diffractive optical element and one focusing lens is created in this work. It can produce rectangular spots, of which the number, the sizes, and the positions are accurately determined by the incident wavelengths. Demonstrated in the customized microflow cytometer, the coefficient of variations (CV) of the optical signals by the beam shaper are 3.6-6.5%, comparable to those derived from the commercial instrument with 3.3-6.3% CVs. Benefiting from the narrow rectangular spots and the flexibility of diffractively shaped lasers, the measurement of bead sizes with 4-15 μm diameters and the real-time detection of flow velocity from 0.79 to 9.50 m/s with the CV of <5% are achieved. © 2020 International Society for Advancement of Cytometry.