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Abstract Active microfluidics for the applications to cellular deformability measurements is an emerging research field ranging from engineering to medicine. Here, we review conventional and microfluidic methods, and introduce an on-chip cell manipulation system with the design principle of fast and fine cell manipulation inside a microchannel. In the latter part of the review, we focus on the results of red blood cell (RBC) deformability measurements as one of the most frequently studied non-adherent cells by on-chip methods. The relationship between mechanical properties and biological structures/features, as well as medical/diagnostic applications, are also discussed.