Differential detachment of intact and viable cells of different sizes using laser-induced microbubbles

Single cell isolation is a prerequisite for the analysis of rare or small cell subtypes. Here, we selectively detach single cells in a heterogeneous population comprised of different morphological subtypes whose sizes vary in body and extension. Such a cellular environment is first accommodated for by a photomechanical method in which pulsed laser irradiation produces microbubbles from a polymer substrate, thus pushing out and detaching cultured cells in an intact, viable, and spatially tailored way. While this has previously only bene used at a very low cell density with lack of quantitative characterization, we determine optimal detachment conditions for different cell sizes in terms of an optical fluence and the number of laser pulses. Importantly, our approach is employed to isolate cancer cells with inherent size variation and elucidate cellular heterogeneity in drug sensitivity: i.e., higher resistance for larger cell size. For cells detached by laser-induced microbubbles, morphology, proliferation, and viability are compared with those of conventional trypsin-treated cells detached without any spatial selectivity. These results support the suitability of our photomechanical method for biochemical screen and secondary analysis of cells with unusual responses.