There are many variations based on the type of tissue to be examined, whether one is dealing with cell suspensions, biopsies, perfused tissues, or monolayer . Fixation Protocol. OSU Campus Microscopy & Imaging Facility :: The Ohio State University College of Med - [Read Fixation Protocol]
Most biological specimens are relatively transparent, so details of internal and intracellular morphology are difficult to image in untreated living specimens using simple bright-field techniques. Fluorescence microscopy offers greater advantages and possibilities for increasing contrast and determining the specific localization of molecules in cells. Article outlines the three methods most commonly used to introduce an appropriate label into Drosophila tissue without perturbing the process. - [Read Fluorescent Reagents for Live Cell Imaging and Their Introduction into Cells]
Frozen Sectioning and Tissue Sectioning for Electron Microscopy. Includes Tissue Processing for Transmission Electron Microscopy and Tissue Preservation for Light Microscopy. University of California UCSF Dept. Pathology Research. - [Read Frozen Sectioning and Tissue Sectioning for Electron Microscopy]
Compendium of protocols for using Aspergillus nidulans in genetic, molecular, and cell biological investigations, originally written for members of my research group. It also summarizes our common growth media and nutritional supplements, many of which originally appeared elsewhere but now are difficult to locate. Includes: Growth and storage of Aspergillus nidulans conidia; Nutritional supplements for our common auxotrophies; Double mutants; Mitotic mapping - assigning genes to chromosomes; etc - [Read Fundamentals of Growth, Storage, Genetics and Microscopy of Aspergillus nidulans Protocols]
In brief, the reaction mixture, is prepared in exactly the same way as if the sample is to be stained or shadowed for electron microscopy, but once the... Gold labeling technique for electron microscopic identification and location of proteins. - [Read Gold labeling of proteins for electron microscopy]
Protocol describes an assay that relies on the endothelial cells’ ability to form distinct blood-vessel-like tubules in an extracellular matrix where they can subsequently be visualized by fluorescence microscopy. Although quantification of the tubules can be performed by manual tracing, this method precludes the use of the assay in unbiased high-throughput applications. - [Read Image Based Assay of Endothelial Cell Tube Formation Protocol]
The scanning transmission electron microscope precision and reproducibility of mass measurements are comparable with those of the analytical ultracentrifuge, the possibility of determining the mass not only of entire supramolecular assemblies but also of their distinct components has opened exciting new avenues which have occasionally been entered but are not yet fully explored. Includes: Principle and application (The GroEL:GroES complex). - [Read Imaging and Measuring Biomolecules & Their Assemblies by Scanning Transmission Electron Microscopy]
Fluorescence microscopy provides a powerful tool for imaging molecular components in living cells. Specific molecular components can be efficiently labeled by a combination of three methods: chemical transfection of GFP-fusion constructs, staining of chromosomes with the DNA-specific, fluorescent dye Hoechst 33342, and microinjection of fluorescently conjugated proteins. This procedure provides an example of using all three methods in sequence to label components of living HeLa cells. - [Read Imaging Hoechst-Labeled Chromosomes and Fluorescent Proteins during the Cell Cycle]
There are several strategies to visualize the antibody. For transmitted light microscopy, color development substrates for enzymes are often used. The antibody can be directly
labeled with the enzyme. However, such a covalent link between an antibody and an enzyme might result in a loss of both enzyme and antibody activity. For these reasons
several multistep staining procedures have been developed, where intermediate link antibodies are used. In this protocol use the Vectastain ABC-kit. - [Read Immunocytochemistry in Free-Floating Sections Protocol]
B or T cells in suspension, adherent cells on chambered coverglass or chamberslides, cryostat sections of unfixed, OCT embedded tissue. Susan Anderson. - [Read Immunofluorescence / confocal microscopy]
Information on: Applications of Confocal Microscopy; Practical Instruments; Limitations of point-scanning Confocal Microscopy; Parallel beam confocal Imaging Systems. - [Read Information on Confocal Imaging]
Article presents an introduction to fluorescence microscopy. Includes: Fundamentals of Excitation and Emission; Stokes' Shift; Fading, Quenching, and Photobleaching; Fluorescence Light Sources; Filter Terminology; The Fluorescence Light Budget; Detecting Single Molecules. - [Read Introduction to Fluorescence Microscopy]
Live-cell imaging techniques provide critical insight into the fundamental nature of cellular & tissue function, especially due to the rapid advances that are currently being witnessed in fluorescent protein & synthetic fluorophore technology. Because of these advances, live-cell imaging has become a requisite analytical tool in most cell biology labs. Includes: Maintaining Live Cells on the Microscope Stage; Live-Cell Imaging Culture Chambers; Optical System and Detector Requirements etc. - [Read Introduction to Live-Cell Imaging Techniques]
Types of light microscopy, Bright Field Microscopy, Using a bright field microscope, mounting on slides, adjusting the microscope, Care of the microscope, When to use bright field microscopy. David R. Caprette. Rice University. - [Read Light Microscopy]
Light Microscopy - Microscopes in Cell Biology. House Ear Institute. Fluorescence microscopy, Nomarski differential interference contrast, Comparison between phase contrast and interference contrast optical systems , Interference contrast, Phase contrast, Darkfield illumination, alignment of Kohler illumination system, Protocol for using oil immersion lenses, Use of immersion oil, Calculating the final magnification on the photomicrograph, vibration, The coverslip glass, Photomicroscopy. - [Read Light Microscopy - Microscopes in Cell Biology]
Ahmanson Center for Advanced Electron Microscopy and Imaging. Use of Semi-Thin Cryosections for Light Microscopy, Immunolabeling of Cryosections on Glass Slides, Problems with Autofluorescence. House Ear Institute - [Read Light Microscopy Techniques]
Protocol for sectioning is on a separate page. 2. Standard Procedure To Section for Transmission Electron Microscopy, EM Center, Indiana University School ... List of standard EM Center procedures/protocols - [Read List of standard EM Center procedures/protocols]