Plant Microtechnique and Microscopy

1. Quick Start Paraffin Embedding Paraffin Embedding - Microwave method Glycol methacrylate embedding Butyl/methyl methacrylate embedding Steedman's wax embedding Chromosome squashes Hematoxylin staining - microwave technique Kohler illumination 2. Microscopy History of the compound microscope Principles of specimen illumination 3. Chemical Fixation of Tissues The quality of fixation The mechanics of fixation Coagulating Fixatives Crosslinking Agents Other fixation methods Recommendations for fixing plant tissues 4. Tissue Dehydration General protocol Dehydration using a graded dehydration series Dehydration using chemical agents 5. Infiltrating Tissues Paraffin infiltrating Paraffin and PEG Embedding Polyethylene glycol and derivatives Plastic infiltrating 6. Sectioning and Mounting Mounting blocks to microtome stubs The quality of sectioning The microtome knife Setting up the microtome Sectioning Methods Mounting sections to glass slides 7. Staining Dye Chemistry Staining equipment General histological staining Other microtechniques stains and protocols Mounting the coverslip Coverslip mounting media 8. Alternate Methods of Microtomy Free-hand Sections Vibratome Cryostat Sliding microtome 9. Special Methods Clearing tissues with NaOH and chloral hydrate Clearing tissues without removing cytoplasmic components Macerating woody tissues Macerating non-woody tissues Bleaching tissues Preserving color in whole mounted specimens 10. Problem Solving Tissue collection Fixation Tannins Hard materials Sliding microtome Clearing Dehydration, Infiltration, and Embedding Microtomy Hand sections Staining 11. Histochemistry and Cytochemistry The relationship of the probe to the target Cytochemical localization of cell components Cytochemical localization of enzymes Cytochemical Fluorescence Microscopy 12. Localization of Molecular Targets Immunolocalization Tissue Printing in situ hybridization APPENDIX I: TOXICS APPENDIX II: BUFFERS APPENDIX III: COMMON CALCULATIONS APPENDIX IV: MERCURY ARC LAMP APPENDIX V: MANUFACTURERS AND VENDORS APPENDIX VI: OPTICS Properties of Light General lens optics The microscope objective The microscope eyepiece The condensor Fluorescence microscopy Microscopy light sources BIBLIOGRAPHY INDEX