Digital holographic microscopy evaluation of dynamic cell response to electroporation
Călin, Violeta L.; Mihăilescu, Mona; Tarbă, Nicolae; Sandu, Ana Maria; Scarlat, Eugen; Moisescu, Mihaela G.; Savopol, Tudor
Biomedical Optics Express
Cell Electroporation (Ep) Consists In Controlled Permeabilization Of The Plasma Membrane By Electric Pulses. Various Molecules (Otherwise Nonpermeant) May Enter Or Exit The Cytoplasm (E.G., Dna, Cytotoxic Drugs, Polysaccharides [1]). Cellular Ep Has Already Proven Valuable In Various Biomedical Fields, As An Adequate Procedure For Drug Delivery (Electrochemotherapy – Standard Clinical Procedure For Treating Tumors [2]), For Gene Electrotransfer (Vaccines, Gene Therapies In Oncology [3]) And For Cell Fusion With Many Biotechnological Applications [4]. Despite These Various Applications, The Dynamics Of The Cellular Response To Electric Pulses Is Only Partially Explored And Our Understanding Is Still Limited. The Experimental Characterization Of The Cell Membrane Reorganization Subsequent To Pulses Application Is Classically Done By Three Categories
Phase-Derived Parameters And Time Autocorrelation Functions Were Used To Analyze The Behavior Of Murine B16 Cells Exposed To Different Amplitudes Of Electroporation Pulses. Cells Were Observed Using An Off-Axis Digital Holographic Microscope Equipped With A Fast Camera. Series Of Quantitative Phase Images Of Cells Were Reconstructed And Further Processed Using Matlab Codes. Projected Area, Dry Mass Density, And Entropy Proved To Be Predictors For Permeabilized Cells That Swell Or Collapse. Autocorrelation Functions Of Phase Fluctuations In Different Regions Of The Cell Showed A Good Correlation With The Local Effectiveness Of Permeabilization.
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