Confocal Microscopy And Multiphoton Excitation Microscopy: by Barry R. Masters

By Barry R. Masters

This article publications you thru the rules and sensible options of confocal and multiphoton microscopy. It additionally describes the ancient connections and parallel innovations that ended in glossy recommendations of reside mobilephone imaging and their use in biology and drugs. you'll find comparisons of other varieties of confocal and multiphoton microscopes, suggestions to the issues one could come upon whilst utilizing numerous microscopic strategies, tips about opting for gear, and an intensive annotated bibliography of extra resources.

Contents

- record of Abbreviations

- Preface

- Optical Microscopy

- Confocal Microscopy

- Nonlinear Microscopy

- the trail to Imaging reside Cells, Tissues, and Organs

- Epilogue

- Appendix: Reference fabrics and assets

- Index

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Extra resources for Confocal Microscopy And Multiphoton Excitation Microscopy: The Genesis of Live Cell Imaging

Sample text

Image brightness in transmission light microscopy is proportional to the square of the ratio of the NA to the total magnification. For the epi-illumination fluorescence mode, the image brightness is proportional to the fourth power of the NA divided by the square of the magnification. Third, the higher the NA, the less the depth of focus. The depth of focus (DOF) is proportional to 1 divided by the square of the NA. Note that the depth of focus is different from the free working distance. The free working distance places a critical limitation on the depth that the objective can focus through the specimen before the specimen makes contact with the tip of the objective.

With Köhler illumination, the iris is located in a plane conjugate to the back focal plane of the objective lens. This iris continuously alters the numerical aperture of the condenser. , using lenses. Catoptric describes an optical system that operates by reflection. Catadioptric refers to an optical system with both reflecting and refracting surfaces that are used to form the image. Two common terms used with lenses are back and front focal planes. The back focal plane of a lens is the focal plane that lies behind the lens when viewed in the direction of the passage of light.

4 Schematic diagram of an epi-illumination fluorescence microscope. A condenser iris diaphragm is located at the front focal plane of the condenser lens of a microscope. With Köhler illumination, the iris is located in a plane conjugate to the back focal plane of the objective lens. This iris continuously alters the numerical aperture of the condenser. , using lenses. Catoptric describes an optical system that operates by reflection. Catadioptric refers to an optical system with both reflecting and refracting surfaces that are used to form the image.

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