Fura-2 Calcium Imaging Microscopy

Calcium measurements using fura-2 were generated using an epifluorescence inverted microscope equipped with a ×20 fluorite objective (33 (link)). Changes in cytosolic calcium levels were monitored in single cells using excitation light provided by a Xenon arc lamp, the beam passing monochromator at 340 and 380 nm (Cairn Research, UK). The emitted fluorescence light was reflected through a 515-nm long pass filter to a cooled CCD camera (Retiga, QImaging, Canada) and digitized to 12-bit resolution. All imaging data were collected and analyzed using software from Andor (Belfast, UK).

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Ludtmann M.H., Arber C., Bartolome F., de Vicente M., Preza E., Carro E., Houlden H., Gandhi S., Wray S, & Abramov A.Y. (2017). Mutations in valosin-containing protein (VCP) decrease ADP/ATP translocation across the mitochondrial membrane and impair energy metabolism in human neurons. The Journal of Biological Chemistry, 292(21), 8907-8917.

Publication 2017

Xenon arc lamp monochromator

Calcium Cells Cytosolic Fluorescence Fura 2 Light Microscope Xenon

Corresponding Organization : University College London

Other organizations : Biomedical Research Networking Center on Neurodegenerative Diseases, Research Institute Hospital 12 de Octubre, Sobell House

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Variable analysis

independent variables

Excitation light provided by a Xenon arc lamp, the beam passing monochromator at 340 and 380 nm

dependent variables

Changes in cytosolic calcium levels

control variables

Epifluorescence inverted microscope equipped with a ×20 fluorite objective
Emitted fluorescence light reflected through a 515-nm long pass filter to a cooled CCD camera
Imaging data collected and analyzed using software from Andor

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