Hostname: page-component-78c5997874-fbnjt Total loading time: 0 Render date: 2024-11-05T08:23:03.310Z Has data issue: false hasContentIssue false

Confocal Microscopy System Performance: Laser Power Measurements

Published online by Cambridge University Press:  14 March 2018

Robert M. Zucker*
Affiliation:
U.S. Environmental Protection Agency, ORD, NHEERL

Extract

Core share and HTML view are not available for this content. However, as you have access to this content, a full PDF is available via the ‘Save PDF’ action button.

The reliability of the confocai laser-scanning microscope (CLSM) to obtain intensity measurements and quantify fluorescence data is dependent on using a correctly aligned machine that contains a stable laser power. The laser power test appears to be one of the most useful tests to quickly evaluate if a system is misaligned or functioning sub optimally by recording insufficient laser power readings. The test using a power meter can indicate if the system is aligned properly up to the plane of excitation on the stage, or if the machine has a defective component (i.e. a dying laser, or a defective fiber). In our experience, without sufficient power throughput in the system, the PMT voltages will have to be increased to high values to visualize fluorescence derived from specimens, which will introduce reduced image quality. In addition the cause of the decreased laser power may result in other problems i.e. laser instability, loss of axial resolution, and increased noise.

Type
Research Article
Copyright
Copyright © Microscopy Society of America 2002

References

References:

1. Zucker, R.M. Price OT Evaluation of confocal system performance. Cytometry 44:273294 2001.Google Scholar
2. Zucker, R.M. Price OT Statistical evaluation of confocal microscopy images. Cytometry 44:295308 2001 3.0.CO;2-C>CrossRefGoogle ScholarPubMed
3. Carter, D: Practical considerations for collecting confocal images. Methods Mol Biol. Confocal Microscopy Methods and Protocols edited by Paddock, S. Humana Press, Totowa NJ. 122:3557 1999.Google Scholar