By David G. Lambert
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Additional resources for Calcium Signaling Protocols (Methods in Molecular Biology Vol 114)
A major limitation of conventional imaging has been that even the high numerical aperture objectives that are used to gain sufficient light for detection also collect light from out-of-focus planes. This has a blurring effect on the resulting image. Confocal microscopy avoids this problem by exciting the indicator and collecting the emission via a pinhole or sometimes a narrow slit (64). The geometry is such that light originating from an out-of-focus plane cannot pass through the pinhole. To construct an image, the ''confocal spot" has to be scanned over the object in view.
Calcium Signaling Protocols results from a chance discussion with Dr. R. I. Norman of the Department of Medicine at Leicester University and represents a major effort from a group of extremely helpful and very patient authors. Putting a book like this together takes time and I am indebted to these authors without whom this project would have remained a chance discussion. I am also very grateful to Professor J. M. Walker, the series editor, for all his help and advice over the course of this project and particularly his help editing the first batch of chapters.
Another indicator in this class is mag-indo-1. It was originally designed for monitoring Mg2+; however, because Mg2+ generally changes very little, these indicators have been used as lowaffinity Ca2+ indicators (Table 3). The dual emission indicators are ideal for simple photometric measurements of Ca2+ from cells. They need only a monochromatic light source (which could be via an interference filter) and a beamsplitting dichroic mirror on the emission side to separate the emission signals (400 and 450 nm for indo-1).
Calcium Signaling Protocols (Methods in Molecular Biology Vol 114) by David G. Lambert