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Infrared microspectroscopy of living cells in microfluidic devices (MD-IRMS)

A novel label-free cell-based assay can be established joining the diagnostic capabilities of IRMS with the design flexibility of microfluidic devices offered by microfabrication. MD-IRMS provides more reliable information on cellular biochemistry in comparison with microspectroscopy on fixed samples and it guarantees the only way to monitor live cells in real time for shining a light on biological events, both naturally occurring or stimulated by mechanical stresses or uptake of chemicals. 


L. Vaccari et al., Anal. Chem. 84(11), 4768 (2012)



Label-free analytical methodologies for studying living cells represent the new frontier of modern cellular biology. As a matter of fact, exogenous probes can bias the cellular response and fixation inevitably alters cellular morphology and biochemistry as well, allowing only the analysis of “static cellular frames”. By combining the diagnostic capabilities of IRMS with the advantages offered by microtechnologies in the design and fabrication of microfluidic devices, we highlighted the biological relevance of MD-IRMS investigating the distinctive biochemical features of live U937 leukemic monocytes in comparison with fixed ones. We established that ethanol fixation dramatically alters both lipid  membrane composition and induces protein aggregation and precipitation while both formalin fixation and air drying only slightly affect lipid and protein cellular pattern. However, any drying procedure induces nucleic acid denaturation through fragmentation and partial transition of the natural B-form of DNA toward the A-form. Therefore, only MD-IRMS can provide reliable information on both cellular DNA and RNA and their modifications in time.

Retrieve Article

Infrared Microspectroscopy of Live Cells in Microfluidic Devices (MD-IRMS): Toward a Powerful Label-Free Cell-Based Assay;
L. Vaccari et al.
Anal. Chem. 84(11), 4768 (2012)

 

 

Last Updated on Friday, 21 December 2012 14:44