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Polarization Modulation Differential Interference Contrast (Pol Mod Dic) Microscopy: An Improvement for Video Microscopy

Published online by Cambridge University Press:  02 July 2020

N. Stromgren Allen
Affiliation:
Department of Botany, Box 7612, North Carolina State University, Raleigh, NC, 27695
D. Moxley
Affiliation:
Department of Botany, Box 7612, North Carolina State University, Raleigh, NC, 27695
D. Collings
Affiliation:
Department of Botany, Box 7612, North Carolina State University, Raleigh, NC, 27695
G. Holzwarth
Affiliation:
Department of Physics, Box 7507, Wake Forest University, Winston-Salem, NC, 27109
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Differential interference contrast (DIC) light microscopy, particularly when coupled with digital image processing, is a powerful tool for the high-resolution microscopy of unstained, transparent biological specimens and can equally well be applied to semiconductor measurements. We show analytically, and with images of diatoms, plant cells and protoplasts, that switching the polarization of the incident light by 90 degrees, changes the image highlights found in conventional DIC images into shadows and vice versa (1). Using a ferroelectric liquid-crystal modulator, this switching can be done at frame rates, synchronized to the camera. By subtracting alternate frames, a stream of difference DIC images is created. We call this technique Pol Mod DIC. Subtraction of alternate images is carried out efficiently by frame buffer operations and amounts to massively parallel synchronous detection. A similar method has been applied to confocal microscopy (2).

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

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References

1.Holzwarth, G. et al. Journal of Microscopy 188 (1997)249.CrossRefGoogle Scholar
2.Corle, T. R. and Kino, G. S.. Applied Optics 29 (1990) 3769.CrossRefGoogle Scholar
3.Allen, R. D. and Allen, N. S.. Journal of Microscopy 129 (1983)3.CrossRefGoogle Scholar
4. This research was supported by NSF facilities grants DIR 87-22684 (to WFU) and BIR-9418205 and North Carolina Biotechnology Center grant 9410-DG-1012 (both to NCSU) and North Carolina Agricultural Experiment Station grant to NSA. We are grateful to Hamamatsu Photonics Inc. and Dage/MTI for making prototype processors available for this study.Google Scholar