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Chapter 33
Visualizing Cells, Tissues, and Molecules

Imaging Biological Samples with Optical Microscopy
Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical…
Phase Contrast and Differential Interference Contrast Microscopy
In-phase-contrast microscopes, interference between light directly passing through a cell and light refracted by cellular components is used to…
Fixation and Sectioning
Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens. The simplest type of…
Immunofluorescence Microscopy
A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy…
Immunocytochemistry and Immunohistochemistry
Immunocytochemistry (ICC) and immunohistochemistry (IHC) are techniques that use antibodies to check for specific proteins or antigens in a sample.…
Confocal Fluorescence Microscopy
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its…
Protein Dynamics in Living Cells
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and…
Total Internal Reflection Fluorescence Microscopy
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a…
Atomic Force Microscopy
Atomic force microscopy (AFM) is a type of scanning probe microscopy that can analyze topographic details of various specimens like ceramics, glass,…
Super-resolution Fluorescence Microscopy
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread…
Overview of Electron Microscopy
The wavelengths of visible light ultimately limit the maximum theoretical resolution of images created by light microscopes. Most light microscopes…
Scanning Electron Microscopy
A scanning electron microscope (SEM) is used to study the surface features of a sample by using an electron beam that scans the sample surface in a…
Transmission Electron Microscopy
In 1931, physicist Ernst Ruska—building on the idea that magnetic fields can direct an electron beam just as lenses can direct a beam of light…
Preparation of Samples for Electron Microscopy
To be visualized by an electron microscope, either transmission or scanning, biological samples need to be fixed (stabilized) so the electron beam…
Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells…
Cryo-electron Microscopy
Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of…
Electron Microscope Tomography and Single-particle Reconstruction
Transmission electron microscopy (TEM) can be used to determine the 3D structure of biological samples with the help of techniques such as electron…
Sequential Immunofluorescence and Immunohistochemistry on Cryosectioned Zebrafish Embryos
Investigation of intercellular interactions often requires discrete labeling of specific cell populations and precise protein localization. The…
Force Spectroscopy of Single Protein Molecules Using an Atomic Force Microscope
The determination of the folding process of proteins from their amino acid sequence to their native 3D structure is an important problem in biology.…
Single Particle Cryo-Electron Microscopy: From Sample to Structure
Cryo-electron microscopy (cryoEM) is a powerful technique for structure determination of macromolecular complexes, via single particle analysis…

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