Fluorescence-based multifunctional light sheet imaging flow cytometry for high-throughput optical interrogation of live cells

Multifunctional light sheet imaging flow cytometry of a large population of live cells at high throughput is challenging and requires new technological advancement. Existing cytometry techniques are limited due to point-based illumination that does not allow volume interrogation and biophysical parameter estimation on the go. Here, we propose a multifunctional (multichannel, multisheet and multicolor) imaging cytometry (M3IC) system that employs vertically-aligned multi-sheet array (VAMSA) illumination for interrogating cells flowing simultaneously through multiple microfluidic channels. We studied cancer cells (volume interrogation with organelle-level resolution and high signal-to-background-ratio(SBR)) at high throughput (similar to 2500 nl/min). M3IC system demonstrates organelle-level resolution with a SBR comparable to that of confocal, especially at low flow rates. In addition, the multicolor imaging capability of the system facilitates multi-organelle investigation, determination of critical biophysical parameters, and drug (Paclitaxel) treatment studies on cancer cells. M3IC system is expected to advance the field of fluorescence microscopy, cell biophysics, disease biology and optical physics.