Non-Destructive On-Chip Imaging Cytometry Assay for Constructive On-Chip Cellomics Studies
from: Flow Cytometry in Microbiology: Technology and Applications (Edited by: Martin G. Wilkinson). Caister Academic Press, U.K. (2015) Pages: 17-58.
We have developed two series of the novel non-destructive imaging cell cytometry assays exploiting microfabrication technology. The on-chip imaging flow cytometry system uses a 1/10,000 s ultra high speed camera with a real-time image analysis unit and a poly(methyl methacrylate (PMMA) based disposable microfluidic chip with the following features: (1) a fully automated system for application and collection of samples using 3-D micropipetting; (2) arrangement of the entire fluidic system in a disposable plastic chip for lining-up of biological samples in laminar flow using hydrodynamic focusing; (3) an optical system for direct observation-based cell identification with specific image indexes using phase-contrast/fluorescence microscopy and real-time image processing; (4) a non-destructive wider-size dynamic range sorting procedure using mild electrostatic force in a laminar flow using agarose gel electrodes to prevent loss of electrode or electrolysis bubble formation; (5) a contamination-free re-cultivation micro-reservoir for collected target cells, and (6) removable DNA aptamer labelling. On-chip single cell imaging cytometry has the advantage of being able to measure the ‘algebraic' inheritance of epigenetic character of single cells using imaging indexes. Exploiting these advantages, target cells can be successfully separated and evaluated and their characteristics by imaging. Results have shown that the on-chip imaging flow cytometry system and the on-chip single-cell imaging cytometry system are applicable for biological research and clinical diagnostics read more ...