High time resolution fluorescence imaging with a CCD camera

• Published in: Journal of neuroscience methods Vol. 36; no. 2-3; p. 253
• Main Authors: Lasser-Ross, N, Miyakawa, H, Lev-Ram, V, Young, S R, Ross, W N
• Format: Journal Article
• Language: English
• Published: Netherlands 01.02.1991
• Subjects: Animals; Calcium - metabolism; Cerebellum - anatomy & histology; Cerebellum - metabolism; Dendrites - ultrastructure; Diagnostic Imaging - instrumentation; Electrodes; Fluorescence; Fura-2; Ganglia, Spinal - ultrastructure; Leeches; Membrane Potentials - physiology; Photography; Purkinje Cells - metabolism
• ISSN: 0165-0270
• DOI: 10.1016/0165-0270(91)90051-Z

We have built a high speed, sensitive camera system capable of capturing sequences of low-light level images synchronized with recordings of membrane potential. The camera system is based on a cooled, scientific grade CCD camera controlled by a PC/AT computer. It can take 100 frames/sec of 18 X 18 element images and 40 frames/sec of 50 X 50 element images with no lag in response to step changes in light intensity. High accuracy and dynamic range of the measurements result from the fact that light levels of the picture elements are digitized with 12 bit accuracy with intrinsic camera noise levels typically less than 1/10,000 of the maximum detectable light level. We have used this system to record calcium dependent fura-2 fluorescence transients in the dendrites of cerebellar Purkinje cells and from different regions of leech neurons in segmental ganglia or isolated in culture.