Fluorescence Microscopy: Super-Resolution and other Novel Techniques delivers a comprehensive review of current advances in fluorescence microscopy methods as applied to biological and biomedical science. With contributions selected for clarity, utility, and reproducibility, the work provides practical tools for investigating these ground-breaking developments. Emphasizing super-resolution techniques, light sheet microscopy, sample preparation, new labels, and analysis techniques, this work keeps pace with the innovative technical advances that are increasingly vital to biological and biomedical researchers.
With its extensive graphics, inter-method comparisons, and tricks and approaches not revealed in primary publications, Fluorescence Microscopy encourages readers to both understand these methods, and to adapt them to other systems. It also offers instruction on the best visualization to derive quantitative information about cell biological structure and function, delivering crucial guidance on best practices in related laboratory research.
- Presents a timely and comprehensive review of novel techniques in fluorescence imaging as applied to biological and biomedical research
- Offers insight into common challenges in implementing techniques, as well as effective solutions
- Provides video accompaniment for certain techniques on a companion website for a multi-dimensional perspective: http://booksite.elsevier.com/9780124095137
Fluorescence Microscopy: Super-Resolution and other Novel Techniques delivers a comprehensive review of current advances in fluorescence microscopy methods as applied to biological and biomedical science. With contributions selected for clarity, utility, and reproducibility, the work provides practical tools for investigating these ground-breaking developments. Emphasizing super-resolution techniques, light sheet microscopy, sample preparation, new labels, and analysis techniques, this work keeps pace with the innovative technical advances that are increasingly vital to biological and biomedical researchers. With its extensive graphics, inter-method comparisons, and tricks and approaches not revealed in primary publications, Fluorescence Microscopy encourages readers to both understand these methods, and to adapt them to other systems. It also offers instruction on the best visualization to derive quantitative information about cell biological structure and function, delivering crucial guidance on best practices in related laboratory research. Presents a timely and comprehensive review of novel techniques in fluorescence imaging as applied to biological and biomedical research Offers insight into common challenges in implementing techniques, as well as effective solutions
Evanescent Excitation and Emission
Abstract
Evanescent light—light that does not propagate but instead decays in intensity over a subwavelength distance—plays a role in fluorescence microscopy in both excitation (i.e., total internal reflection, or TIR) and emission (i.e., supercritical angle fluorescence). This chapter describes the physical connection between these two forms as a consequence of geometrical compression of wavefront spacing and describes newly established or speculative applications and combinations of the two. In particular, each form can be used in analogous ways to produce surface-selective images, to examine the thickness and refractive index of films (such as lipid multilayers or protein layers) on solid supports, and to measure the absolute distance of a fluorophore to a surface. In combination, the two forms can further increase selectivity and reduce background scattering in surface images. The polarization properties of each lead to more sensitive and accurate measures of fluorophore orientation and membrane micromorphology. The phase properties of evanescent excitation lead to methods of creating a submicroscopic area of TIR illumination or enhanced-resolution structured illumination. Analogously, the phase properties of evanescent emission lead to a method of producing a smaller point spread function, in a technique called virtual supercritical angle fluorescence. This chapter emphasizes the concepts and theory (rather than experimental protocols and results) of evanescence for both excitation and emission, as well as the theory of its many existing and possible future applications.
Keywords
microscope imagingnear fieldpoint spread functionpolarizationsupercritical anglesuper-resolutiontotal internal reflection
Introduction
Evanescence in General
(1.1)
The orientation of the (x, y, z) axes can be arbitrary but is chosen based on the geometry of optical surfaces nearby. The amplitude of km, called the wavenumber, is(1.2)
where ω is the angular frequency of the particular color of the light, and c is the speed of light in vacuum. Frequency ω is equal everywhere in the optical system, regardless of refractive index.(1.3)
The position-dependent part of the plane wave electric field has the form exp(ikm·r), where position vector . Therefore, the spatial variation of the electric field in, say, the z direction is the sinusoidal function exp(ikmzz).Erscheint lt. Verlag | 24.2.2014 |
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Sprache | englisch |
Themenwelt | Naturwissenschaften ► Biologie ► Biochemie |
Naturwissenschaften ► Biologie ► Genetik / Molekularbiologie | |
Naturwissenschaften ► Biologie ► Zellbiologie | |
Technik | |
ISBN-10 | 0-12-416713-6 / 0124167136 |
ISBN-13 | 978-0-12-416713-1 / 9780124167131 |
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