Regensburg 2022 – wissenschaftliches Programm
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BP: Fachverband Biologische Physik
BP 12: Poster 2
BP 12.49: Poster
Dienstag, 6. September 2022, 17:30–19:30, P4
Establishment of a microfluidic UV-Vis analysis of single cell E. coli — •Tim R. Baumann1, Alexander Grünberger2, Dario Anselmetti1, Harald Gröger3, and Martina Viefhues1 — 1Experimental Biophysics & Applied Nanosciences, Department of Physics, Bielefeld University — 2Multiscale Bioengineering, Department of Technology, Bielefeld University — 3Industrial organic chemistry and biotechnology, Department of Chemistry, Bielefeld University
Whole-cell biocatalysts like, E. coli DH5-α are widely used in industrial organic chemistry and biotechnology. For efficient production an appropriate cultivation media composition is of high importance. Harmful organic solvents, like ethanol are needed to solve insoluble substrates, those often lead to a permeabilisation of the bacterial membranes. In this study, we established a microfluidic method to analyse the impact of ethanol on leaching of certain intracellular cofactors e.g., NAD(P)H and exploited the contribution of NAD(P)H to the cells autofluorescence. In order to detect the losses in intensity due to leaching, a UV-LiF analysis of single cells was conducted, using a Nd:YAG Laser (λ = 266 nm). We evaluated the impact of incubation in 5 % ethanol for either 5 or 10 minutes on single cells in a PDMS microfluidic chip with a UV transparent fused silica base layer, including a carbon black PDMS spot at the detection point to reduce the PDMS’s autofluorescence. The intensity data was assessed and plotted in histograms. Those exhibited a reduction of the FWHM and a displacement of the distribution maxima to smaller intensities depending on the incubation time and proved the cofactor leaching due to ethanol exposition.