MCDB 423 Lecture Notes - Lecture 11: Nature Neuroscience, Design Of Experiments, Oregano
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23 Dec 2020
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In notional cell calcium is a essential messenger. Calcium green: single wavelength calcium indicator (no change when binding, ex/em: calcium green 506/531 nm. Fluorescence intensity increase upon binding ca2+ : ~14 fold: once you stimulate ion channel calcium rushes into cell and since pre loaded with calcium green that calcium will bind into dye and result in increase in amount of. Fluorescence you can detect: monitoring function of receptor through increased amount of uorescence within the cells transfected ion channel. Transfect hek cells with trpv3 ions channel plasmid 2 days prior to imaging. Incubate cells with calcium green: wash cells, assess trpv3 ion effect on calcium current by treating transfected cells with different solutions/experimental conditions. Experimental design: carvocrol, heat, how fast should we collect our frames (time laps imaging), oregano, thyme and clove-derived avors and skin sensitizers activate speci c trp channels.
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