光学&电子&荧光显微技术

光学显微技术

• The Light Microscope (House Ear Institute)
  An explanation of how the light microscope works, how to use it, and how to get optimal results when using it.
    
• Photomicrography(Bowtell Lab)

• Immunolabeling for light microscopy (House Ear Institute)
  Simple method for preparing thin sections of biological material for light microscopy. These sections give better high resolution information and provide better specimen preservation than even the thinnest cryostat section. May even be better than confocal optical sections!
    
• Mounting medium for Immunofluorescence microscopy (House Ear Institute)
  This mounting medium is easy to prepare, can be stored cold or frozen, can be used with anti-fade agents and will polymerize to give a dry, semi-permanent mount. If for some reason the sample is needed again, immersing in water will dissolve the medium again

电子显微技术

• The Transmission Electron Microscope (TEM) (HEI)
  An explanation of how the TEM works.
    
• TEM Specimen Preparation (HEI)
    
• Serial Sectioning (Walter Steffen)
     
• EM visualization techniques (HEI)
  An introduction to the routine methods used when preparing specimens for transmission electron microscopy.
    
• Negative staining for transmission electron microscopy. (HEI)

o  Introduction.

o  Using coated grids.

o  Using carbon films.

o  Staining protocols.

o  Staining solutions.

• High resolution negative staining (Microscopy and Electronic Imaging Lab)
   
• Preparation of Holey Carbon Film (Walter Steffen)
    
• Negative Staining (Walter Steffen)
    
• Embedding in epoxy resins (HEI)
  Process cultured cell for EM
   
• EPON resin mixture for transmission electron microscopy (Microscopy and Electronic Imaging Lab)
    
• How to coat metal specimen grids with Formusr/local (HEI)
  Strong, thin support films are essential for routine TEM.
  Here is a reproducible method for making thin films of formusr/local with details on how to mount them on metal specimen grids.
   
• Preparation Of Ciliated Protozoa For Scanning Electron Microscopy(Microscopy and Electronic Imaging Lab)
  This procedures is used to fix and stain cells for SEM and for TEM. Cells can be fixed using conventional glutaraldehyde-osmium fixation  To preserve ciliary orientation, use the "instant fixation" protocol described here. With this method, the cortex and ciliary beat form is well preserved but the cytoplasm is poorly preserved and membrane breakage and blebbing on the cell surface is evident.
• Specimen Preparation for Scanning Electron Microscopy(Microscopy and Electronic Imaging Lab)
  Methods for preparation  of a usr/localiety of specimens.
    
• Sample Fixation for Electron Microscopy(Microscopy and Electronic Imaging Lab)
  This method produce good fixation for most tissues.
    
• Chlamydomonas Fixation for Transmission Electron Microscopy (Microscopy and Electronic Imaging Lab)
    
• Fixation of Cells Cultured in Transwell Dishes (Microscopy and Electronic Imaging Lab)
  Transwell culture dishes are commonly used to culture cells so that the top and bottom of the cells can be exposed to different culture media conditions. They are commonly used with polarized epithelial cells and endothelial cells. The cells are cultured on membrane supports and processing for transmission electron microscopy is greatly facilitated if the membranes are left intact and under the tension placed by the wells in the dish. Most dishes are destroyed by solvents used in embedding but the following method, when used with polycarbonate Transwell plates obtained from Costar, works very well.
    
• Fixation and Embedding of Microtubules for Electron Microscopy (Microscopy and Electronic Imaging Lab)
  (This procedure can also be used for virtually any material that must be pelleted prior to fixation and thin sectioning)
• Tetrahymena Fixation for Transmission Electron Microscopy (Microscopy and Electronic Imaging Lab) 
• Preparation of Yease Cell for ImmunoEletron Microscopy (Jon Mulholland)

荧光显微技术

• Immunofluorescencc Microscopy of tissue culture cells (Microscopy and Electronic Imaging Lab)
  These methods are written for direct staining of filamentous actin with bodipy FL-phallicidin and indirect immunofluorescence staining of microtubules with anti-tubulin antibodies. They easily can be modified for conventional immunofluorescence staining of any tissue cultured cell or direct staining of actin filaments
   
• Photomicrography(Bowtell Lab)
     
• Staining Nucleic Acids with Propidium Iodide (Molecular Dynamics Inc)
  Propidium iodide is a membrane-impermeant dye that stains by intercalating into nucleic acid molecules. It binds both DNA and RNA.It is useful in confocal microscopy studies of chromosome topography, DNA structure, and membrane integrity. It can also be used in double-label experiments to detect DNA and a fluorescein-labeled antigen simultaneously. This protocol can be used to stain cells with propidium iodide alone, simultaneously with another fluorochrome, or after staining with another fluorochrome.
    
• Immunfluoresence on Chamber Slides (Bowtell Lab)
   
• Mounting medium for immunofluorescence microscopy (House Ear Institute)
  This mounting medium is easy to prepare, can be stored cold or frozen, can be used with anti-fade agents and will polymerize to give a dry, semi-permanent mount. If for some reason the sample is needed again, immesing in water will dissolve the medium again.