To the best of my knowledge, biocytin can be used to identify the patched cell but only after the histology staining. As I would like to label/mark the living cell that have been patched, I was wondering whether there is a technique/dye that would allow me to stain the cell alive but with least side effects or toxicity to the cell. So that I could visualize the cell under bright-field microscope and do my patch clamping.
As I am not going to use fluorescence, thus GFP injection technique is not useful to me.
By the way, thank you very much in advance.
A very common way to visualize a patch-clamped cell is to include a fluorophore in the intra-cellular solution. For example adding 100 uM Alexa 488 that emits green when excited by blue light (around 488nm, hence the name). After going whole-cell, the fluorophore will distribute in the whole cell by diffusion, making it easily recognisable. You will need an excitation light source, and appropriate excitation and emission filters to see the fluorescence, though this is quite standard equipment in most patch-clamp rigs.
Actually, I am thinking of the medical grade inks, which suppose to possess least toxic side effect to the body while being visible under normal illumination. I was wondering whether anyone may provide more cytotoxicity information about these inks.
Thank you very much in advance.
Biocytin 0.1 - 0.5 % in your internal. Stain that up how you like. Works a treat.
Alexa and Biocytin are the most common and fairly straight forward.....
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