I'm looking for either advice or maybe someone who has previously done calcium imaging with organotypic brain cultures. I'm looking to monitor the intracellular calcium levels while stimulating the culture or at least get an idea of how this stimulation changed the calcium levels from normal. I culture my brain slices on top of a millicell insert membrane. I have read that certain calcium-binding dyes have been known to attach to the membrane strongly, so I'm looking for possible dye suggestions and possible procedures for accomplishing intracellular calcium monitoring. Any help would be greatly appriciated!!!