High-throughput imaging flow cytometry by optofluidic time-stretch microscopy

Cheng Lei, Hirofumi Kobayashi, Yi Wu, Ming Li, Akihiro Isozaki, Atsushi Yasumoto, Hideharu Mikami, Takuro Ito, Nao Nitta, Takeaki Sugimura, Makoto Yamada, Yutaka Yatomi, Dino Di Carlo, Yasuyuki Ozeki, Keisuke Goda

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88 Citations (Scopus)


The ability to rapidly assay morphological and intracellular molecular variations within large heterogeneous populations of cells is essential for understanding and exploiting cellular heterogeneity. Optofluidic time-stretch microscopy is a powerful method for meeting this goal, as it enables high-throughput imaging flow cytometry for large-scale single-cell analysis of various cell types ranging from human blood to algae, enabling a unique class of biological, medical, pharmaceutical, and green energy applications. Here, we describe how to perform high-throughput imaging flow cytometry by optofluidic time-stretch microscopy. Specifically, this protocol provides step-by-step instructions on how to build an optical time-stretch microscope and a cell-focusing microfluidic device for optofluidic time-stretch microscopy, use it for high-throughput single-cell image acquisition with sub-micrometer resolution at >10,000 cells per s, conduct image construction and enhancement, perform image analysis for large-scale single-cell analysis, and use computational tools such as compressive sensing and machine learning for handling the cellular ‘big data’. Assuming all components are readily available, a research team of three to four members with an intermediate level of experience with optics, electronics, microfluidics, digital signal processing, and sample preparation can complete this protocol in a time frame of 1 month.
Original languageEnglish
Pages (from-to)1603-1631
Number of pages29
JournalNature Protocols
Issue number7
Publication statusPublished - 5 Jul 2018

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