Chapter 8
Microscopy
Abstract Many technical developments have appeared in recent years to enhance
imaging performance and instrument versatility in optical microscopy for biopho-
tonics and biomedical applications to visualize objects ranging in size from mil-
limeters to nanometers. These developments include increasing the penetration
depth in scattering media, improving image resolution beyond the diffraction limit,
increasing image acquisition speed, enhancing instrument sensitivity, and devel-
oping better contrast mechanisms. This chapterfirst describes the basic concepts
and principles of optical microscopy and then discusses the limitations of distin-
guishing two closely spaced points. Next, the functions of confocal microscopes,
fluorescence microscopy, multiphoton microscopy, Raman microscopy, light sheet
microscopy, and super-resolutionfluorescence microscopy are given.
Microscopy applications can be found in all branches of science and engineering.
The diverse instruments used in thesefields include acoustic microscopes, atomic
force microscopes, scanning electron microscopes, x-ray microscopes, and optical
microscopes. This chapter addresses optical microscopes and their applications to
biophotonics. Many technical developments have appeared in recent years to
enhance imaging performance and instrument versatility in optical microscopy for
biophotonics and biomedical applications. These developments include increasing
the penetration depth in scattering media, improving image resolution beyond the
diffraction limit, increasing image acquisition speed, enhancing instrument sensi-
tivity, and developing better contrast mechanisms. Different embodiments of these
optical microscopic techniques can visualize objects ranging in size from mil-
limeters to nanometers. Microscopes can be categorized by their physical structure,
functional principle, illumination method, and image processing procedure.
Applications of microscopy instruments and techniques are found in all branches of
biomedicine and biophotonics, for example, analyses of biological samples, brain
research, cancer research, cytology, drug discovery, food analyses, gynecology,
healthcare, hematology, and pathology.
First Sect.8.1describes the basic concepts and principles of optical microscopy.
This is followed in Sect.8.2by discussions of the limitations of distinguishing two
©Springer Science+Business Media Singapore 2016
G. Keiser,Biophotonics, Graduate Texts in Physics,
DOI 10.1007/978-981-10-0945-7_8
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