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Fluorescent Speckle Microscopy

Published online by Cambridge University Press:  14 March 2018

Stephen W. Carmichael*
Affiliation:
Mayo Clinic
Wilma L. Lingle
Affiliation:
Mayo Clinic

Extract

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Whereas too much of a good thing can be bad, too little of a good tiling can be good. In this case, the accidental dilution of X-rhodamine tubulin injected into living cells resulted in a heterogeneous labeling of micro tubules, rather than visualization of continuous structures. This heterogeneous labeling rendered a speckled image, hence the term fluorescent speckle microscopy (FSM) was introduced. It turns out that FSM yields particularly useful information on dynamic events within living cells, for example the assembly and disassembly of microtubules.

There are two recent reviews of FSM by Clare Waterman-Storer and Gaudenz Danuser, one emphasizing the biologic applications of the technique, the other emphasizing the quantitative aspects. For their purposes, they defined a “speckle” as a diffraction-limited region of the image that is significantly brighter than its immediate environment. It can be calculated from the point-spread function (which is determined by the numerical aperture of the lens) that optimally a diffraction-limited region of about 250 nm can be imaged.

Type
Research Article
Copyright
Copyright © Microscopy Society of America 2004

Footnotes

1

The authors gratefully acknowledge Dr. Clare Waterman-Storer for reviewing this article.

References

2 Waterman-Storer, C.C., and Danuser, G., New directions for fluorescent speckle microscopy, Current Biology 12:R633R640, 2002.Google Scholar
3 Danuser, G., and Waterman-Storer, C.M., Quantitative fluorescent speckle microscopy: Where it came from and where it is going, J. Microscopy 211:191207, 2003.Google Scholar