In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons

Kalb J, Nielsen T, Fricke M, Egelhaaf M, Kurtz R (2004)
Biochemical and biophysical research communications 316(2): 341-347.

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Abstract
We applied two-photon laser-scanning microscopy (TPLSM) to motion-sensitive visual interneurons of the fly to study Ca(2+) dynamics in vivo at a higher spatial and temporal resolution than possible with conventional fluorescence microscopy. Based on a custom-built two-photon microscope, we performed line scans to measure changes in presynaptic Ca(2+) concentrations elicited by visual stimulation. We used a fast avalanche photodiode (APD) with a high quantum efficiency to detect even low levels of emitted fluorescence. Our experiments show that our in vivo preparation is amenable to TPLSM: with excitation intensities low enough not to cause photodamage, activity-dependent fluorescence changes of Ca(2+)-sensitive dyes can be detected in small neuronal branches. The performance of two-photon and conventional Ca(2+) imaging carried out consecutively at the same neuron is compared and it is demonstrated that two-photon imaging allows us to detect differences in Ca(2+) dynamics between individual neurites.
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Kalb J, Nielsen T, Fricke M, Egelhaaf M, Kurtz R. In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons. Biochemical and biophysical research communications. 2004;316(2):341-347.
Kalb, J., Nielsen, T., Fricke, M., Egelhaaf, M., & Kurtz, R. (2004). In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons. Biochemical and biophysical research communications, 316(2), 341-347.
Kalb, J., Nielsen, T., Fricke, M., Egelhaaf, M., and Kurtz, R. (2004). In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons. Biochemical and biophysical research communications 316, 341-347.
Kalb, J., et al., 2004. In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons. Biochemical and biophysical research communications, 316(2), p 341-347.
J. Kalb, et al., “In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons”, Biochemical and biophysical research communications, vol. 316, 2004, pp. 341-347.
Kalb, J., Nielsen, T., Fricke, M., Egelhaaf, M., Kurtz, R.: In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons. Biochemical and biophysical research communications. 316, 341-347 (2004).
Kalb, Julia, Nielsen, Tim, Fricke, Matthias, Egelhaaf, Martin, and Kurtz, Rafael. “In vivo two-photon laser-scanning microscopy of Ca 2+ dynamics in visual motion-sensitive neurons”. Biochemical and biophysical research communications 316.2 (2004): 341-347.
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