Figure 4
Targeted laser-assisted transfection of multiple opsins: functional evaluation and quantitative comparison between two non-viral delivery methods. (a) Fluorescence image of HEK293 cells after optoporation with White-opsin, imaged using a 100× objective. Scale bar: 15 µm. (b) Representative fluorescence intensity profiles of reporter proteins along lines drawn across cells (e.g., the dotted white line in a). (c) Quantitative comparison of the expression in the membranes and intracellular components of cells optoporated with White-opsin. N = 5. Average ± S.D. *p < 0.01 difference between the membranes and intracellular components of White-opsin-expressing cells. (d) Representative inward photocurrent profiles in response to white-light optogenetic stimulation (0.12 mW mm−2) at two different pulse widths (100 and 200 ms). (e) Quantitative comparison between the opsin expression distributions (ratios of membrane to intracellular expression) observed after optoporation and lipofection. N = 7/opsin/delivery method. Average ± S.D. *p < 0.05 difference between membrane and intracellular components. Flour., fluorescence.
