Learning Calcium Imaging - What is a good introductory resource for learning how to perform calcium imaging?

Resources
1

I want to add calcium imaging (of dissociated neurons in a dish) to my repertoire of skills. I’m seeking a good introductory learning resource that has very practical info to help me execute my own calcium imaging experiments. I can’t seem to find anything that fits the bill. There are reviews on the conceptual aspects of calcium imaging, but not so much on the specific details of doing calcium imaging (e.g. what equipment, what dyes, which programs to analyze data, etc.).

Any recommendations would be great. Thanks.

Cross-Posted @ ResearchGate

2

We routinely perform Calcium imaging with a Fura-2am dye and use a DG4 as our light source. Once you have the equipment it is fairly straight forward, but it also is fairly expensive to set up with the scope that is required, the filter cubes, DG4 (or Lambda as light source). We just upgraded our second system and are now using NIH elements for both computers as our software, but the analysis is all by hand in an excel chart.
If you have more questions let me know.

3

Thanks @fmoehring. Besides the equipment, did you read any articles/texts or anything to explain the details of setting up your experiments properly, or did you just learn from people in your lab?

4

I was taught how to perform the experiments by our technician but there are papers out there that utilize the technique extensively and which we always use as a reference for what agonist concentrations to use, how long to apply each agogist, etc. I can look through my materials and sent you some of them if you want.

5

That would be great if you’d post some of those key papers here so that myself and others could use. Thanks!

6

I really don’t know how helpful this paper will be for any of you, because it only has the dose response curves for TRPA1 and TRPV1 agonists, but those are the agonists we work with on a daily basis. But the method section describes our protocols pretty well. To test viability of the neurons we always use 50mM K+.

TRPA1 is functionally expressed primarily by IB4-binding, non-peptidergic mouse and rat sensory neurons.
Barabas, Marie E; Kossyreva, Elena A ;Stucky, Cheryl L

7

Has anyone here done calcium imaging of spinal cord slices with either dye or GCaMP? If you have a protocol, would you mind sharing?