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Multi-Mode Light Microscopy of Microtubule and Endoplasmic Reticulum Dynamics in Migrating Newt Epithelial Cells.

Published online by Cambridge University Press:  02 July 2020

C. M. Waterman-Storer  and
E. D. Salmon
C. M. Waterman-Storer
Affiliation:
Department of Biology, University of North Carolina, Chapel Hill, NC, 27599-3280
E. D. Salmon
Affiliation:
Department of Biology, University of North Carolina, Chapel Hill, NC, 27599-3280
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Extract

We have developed a multi-mode digital imaging system (1-3) which acquires images with a 12 bit cooled CCD camera. A multiple band pass dichromatic mirror and robotically controlled excitation filter wheels provide rapid wavelength selection for epi-fluorescence with DAPI, fluorescein or GFP and X-rhodamine fluorophores while maintaining image registration on the cooled CCD detector. Shutters select illumination either by epi-fluorescence or by transmitted light for phase contrast or DIC. A robotically controlled emission filter wheel in front of the CCD camera inserts an analyzer in the light path for DIC imaging. To maximize fluorescence light intensity, the analyzer is removed and an optical flat of equivalent optical thickness is inserted for fluorescence imaging. A slider is inserted at the field diaphragm position of the fluorescence epi-illuminator to provide in-focus slit and spot targets for 360 nm photoactivation of “caged” fluorophores. The microscope system is robotically controlled and image acquisition and analysis is performed using MetaMorph™ digital imaging software.

Type
Innovative Approaches to 3-D Structure/Function Determination for Cells and Organelles
Information
Microscopy and Microanalysis , Volume 3 , Issue S2: Proceedings: Microscopy & Microanalysis '97, Microscopy Society of America 55th Annual Meeting, Microbeam Analysis Society 31st Annual Meeting, Histochemical Society 48th Annual Meeting, Cleveland, Ohio, August 10-14, 1997 , August 1997 , pp. 211 - 212
Copyright
Copyright © Microscopy Society of America 1997

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References

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6. Supported by NIH grant GM24364Google Scholar