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A Transportable Multiple Oblique Illumination System which Retrofits to Conventional Optical Microscopes to Provide Highdefinition Real Time 3-Dimensional Imaging

Published online by Cambridge University Press:  02 July 2020

Gary L. Greenberg
Affiliation:
Edge Scientific Instrument Co., LLC, 1630 17th St., Santa Monica, CA, 90404
Alan Boyde
Affiliation:
Department of Anatomy and Developmental Biology, University CollegeLondon
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Microscope specimens are 3-dimensional objects, but the images from conventional light microscopes are flat - they only show two dimensions. Multiple oblique illumination is a novel lighting technique for transmitted light microscopes that produces multiple (i.e. two or more) high definition 3-dimensional images using conventional microscope objective lenses. We have previously described its use in transmitted light. The same optical theory has now been expanded to include reflection microscopy. In the present paper, we describe a new development which will make this approach more widely available. It is a retrofit illumination system that will produce true 3-dimensional images directly through the eyepieces of conventional microscopes.

By seeing z-axis information in real-time and in the context of a specimen's entire thickness, researchers can gain additional, unambiguous information about the interrelationships between structures, whereas critical information about 3-dimensional structures can be obscured, lost or misinterpreted when using 2-dimensional instruments. The importance of accurate z-axis information has popularized methods such as deconvolution and confocal microscopy.

Type
Unique Approaches in Imaging, Computation and Communication for Characterization of the 3D Cell & Organelles I
Copyright
Copyright © Microscopy Society of America

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References

References:

1.Greenberg, G., Boyde, A.. Novel method for stereo imaging in light microscopy at high magnifications. Neurolmage 1: (1993) 121128.Google Scholar
2.Greenberg, G, Novel methods for direct-view 3D imaging in optical microscopes. Proc roy microsc Soc 31(2): (1996) 150151.Google Scholar
3.Fay, F.S., et. al. Three-dimensional molecular distribution in single cells analyzed using the digital imaging microscope. J. Microsc. 153:133–49 (1989).CrossRefGoogle Scholar
4.Pawley, J.B. (ed) Handbook of Biological Confocal Microscopy, New York: Plenum Press (1995).CrossRefGoogle Scholar
5.Greenberg, G. & Boyde, A.Convenient and controllable direct-view 3D imaging in conventional optical microscopes: Approaches via illumination and inspection. Proc. Roy Mircosc Soc 32/2:87101 (1997a).Google Scholar
6. We both thank S Katayama and R Kasper for their consistent and generous support of the developments described here.Google Scholar