title: Assessment of enhanced autofluorescence and impact on cell microscopy for microfabricated thermoplastic devices.
abstract: Thermoplastics such as polystyrene (PS) and cyclo-olefin polymer (COP) have become common materials for fabrication of microfluidic cell-based systems because of a number of attractive properties. However, thermoplastics are also known to exhibit autofluorescence levels that may hinder their utility for cell-based and imaging applications. Here, we identify and characterize a phenomenon causing an increase in the autofluorescence of polystyrene after thermal treatment. This effect is of particular importance for plastic microfluidic device fabrication because the ranges of pressures and temperatures causing this effect match the same range as those used for polystyrene bonding. Further, we find that the enhanced autofluorescence has significant impact on the image quality, accuracy, and ability to identify and quantify fluorescently labeled cells. We tested two alternative strategies, solvent bonding of PS or thermal bonding of COP, to alleviate the adverse effects of heterogeneous and enhanced autofluorescence on cell image analysis, and demonstrate that both strategies are viable options to thermal bonding of PS for specific applications where cellular imaging is of primary interest.
authors.affiliation: Department of Biomedical Engineering, Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, 53705, United States.
authors.forename: Edmond W K
authors.initials: EW
authors.lastname: Young
authors.forename: Erwin
authors.initials: E
authors.lastname: Berthier
authors.forename: David J
authors.initials: DJ
authors.lastname: Beebe
mesh_terms: D000475:Alkenes / Q000737:chemistry; D018951:Antigens, CD20 / Q000378:metabolism; D045744:Cell Line, Tumor; D005456:Fluorescent Dyes / Q000737:chemistry*; D005464:Fluorobenzenes / Q000737:chemistry; D006801:Humans; D018613:Microscopy, Confocal / Q000295:instrumentation / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D011137:Polystyrenes / Q000737:chemistry; D011312:Pressure; D012997:Solvents / Q000737:chemistry; D053668:Syndecan-1 / Q000378:metabolism; D013696:Temperature
chemical_list: C513915:AlexaFluor 488 carboxylic acid tetrafluorophenyl ester; D000475:Alkenes; D018951:Antigens, CD20; D005456:Fluorescent Dyes; D005464:Fluorobenzenes; D011108:Polymers; D011137:Polystyrenes; D012997:Solvents; D053668:Syndecan-1