Hi all. Wanting to do some calcium imaging, either with Fura-2 or GCaMP on dissociated DRGs. Does anyone have a detailed protocol they use? We have one, but I’m wanting to just cross-reference with others to optimize. @tberta @thicunha @defaria
@achamess my lab is also starting on calcium imaging, and I will be glad to have good protocols. We are planning to use HCS machines (Image express) with a robotic pipette.
For Fura-2, Michael Gold’s lab is very rigorous ( PMID: 23642703), ours is a little more basic but works well: PMID: 20398327. You can combine with GFP or live cell imaging like IB4 or other cell-surface antigens with directly-conjugated antibodies. With good optics you can use 20x and image lots of cells, or get better resolution at 40x. You can also pick up the cells afterwards for single cell PCR if you want to make the effort.
1 Like