Hostname: page-component-7bb8b95d7b-dtkg6 Total loading time: 0 Render date: 2024-09-11T14:32:56.056Z Has data issue: false hasContentIssue false

Multispectral Imaging in Light Microscopy

Published online by Cambridge University Press:  02 July 2020

K. R. Spring*
Affiliation:
Laboratory of Kidney and Electrolyte Metabolism, National Heart, Lung and Blood Institute, NIH, Bethesda, MD, 20892-1603
Get access

Extract

Many recent applications in light microscopy involve the use of multiple fluorophores or the delineation of signals arising from spectrally distinct sources. In microspectroscopy, it is always desirable to illuminate fluorescently-labeled microscopic specimens with monochromatic light as the narrowest possible excitation wavelength range usually results in the highest emission signal-to-noise ratio. Generation of polychromatic light from an arc lamp and selection of the excitation wavelength by interference filters or monochrometers are the most common techniques for excitation microspectrofluorometry. Emission spectroscopy is usually done with filter wheels, monochrometers, or interferometers inserted between the microscope detection port and the detector. This presentation will be directed toward other, less frequently-used, approaches for spectral scanning of the specimen in the light microscope. Three topics will be considered: 1) the use of acousto-optical tunable filters and lasers for rapid, narrow-band, excitation wavelength selection; 2) the use of holographic notch filters for rejection of unwanted excitation laser light; 3) using liquid-crystal tunable filters for emission scanning.

Type
Compositional Mapping With High Spatial Resolution
Copyright
Copyright © Microscopy Society of America

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)