Neuronal Response to Caffeine Using a Chemical Probe in SPM 

Functional Calcium Imaging - An Optical Alternate to Patch Clamping

• In this series of images a Nanopipette probe containing OrangeGreen is held in place with AFM control over a neuronal cell membrane of a cell which is filled with Fluo3.
• The probe tip and the cell are excited in epi-illumination.
• Each frame in the image is one confocal scan with 250 millisecond intervals between frames.  
• A puff of caffeine is injected just before the fourth image (top row).
• The cell and tip respond to the caffeine peaking in the center of the second row. Some of the pixels are red indicating that calcium has flowed out of the membrane and can be found at the tip which is in close proximity to the cell membrane.
• Only Nanonics produces such probes for chemical imaging.