title: Infrared microspectroscopy of live cells in microfluidic devices (MD-IRMS): toward a powerful label-free cell-based assay.
abstract: Until nowadays most infrared microspectroscopy (IRMS) experiments on biological specimens (i.e., tissues or cells) have been routinely carried out on fixed or dried samples in order to circumvent water absorption problems. In this paper, we demonstrate the possibility to widen the range of in-vitro IRMS experiments to vibrational analysis of live cellular samples, thanks to the development of novel biocompatible IR-visible transparent microfluidic devices (MD). In order to highlight the biological relevance of IRMS in MD (MD-IRMS), we performed a systematic exploration of the biochemical alterations induced by different fixation protocols, ethanol 70% and formaldehyde solution 4%, as well as air-drying on U937 leukemic monocytes by comparing their IR vibrational features with the live U937 counterpart. Both fixation and air-drying procedures affected lipid composition and order as well as protein structure at a different extent while they both induced structural alterations in nucleic acids. Therefore, only IRMS of live cells can provide reliable information on both DNA and RNA structure and on their cellular dynamic. In summary, we show that MD-IRMS of live cells is feasible, reliable, and biologically relevant to be recognized as a label-free cell-based assay.
authors.affiliation: Elettra Synchrotron Light Laboratory, SISSI beamline, S.S. 14 Km 163.5, 34149 Basovizza, Trieste, Italy. lisa.vaccari@elettra.trieste.it
authors.forename: L
authors.initials: L
authors.lastname: Vaccari
authors.forename: G
authors.initials: G
authors.lastname: Birarda
authors.forename: L
authors.initials: L
authors.lastname: Businaro
authors.forename: S
authors.initials: S
authors.lastname: Pacor
authors.forename: G
authors.initials: G
authors.lastname: Grenci
mesh_terms: D045744:Cell Line, Tumor; D004247:DNA / Q000032:analysis*; D003890:Desiccation; D000431:Ethanol / Q000737:chemistry; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D006801:Humans; D008055:Lipids / Q000737:chemistry; D046210:Microfluidic Analytical Techniques; D008855:Microscopy, Electron, Scanning; D008968:Molecular Conformation; D009000:Monocytes / Q000737:chemistry* / Q000648:ultrastructure; D011506:Proteins / Q000032:analysis*; D012313:RNA / Q000032:analysis*; D017550:Spectroscopy, Fourier Transform Infrared; D014867:Water / Q000737:chemistry
chemical_list: D005404:Fixatives; D008055:Lipids; D011506:Proteins; D014867:Water; D005557:Formaldehyde; D000431:Ethanol; D012313:RNA; D004247:DNA