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This e-book provides the latest know-how in electron microscopy. It includes significant parts of electron microscopy - transmission electron microscopy (TEM) and scanning electron microscopy (SEM). the quantity presents transparent, concise directions on processing organic specimens and contains dialogue at the underlying rules of the vast majority of the procedures provided. A notes part allows effective edition and troubleshooting of protocols.
Read Online or Download Electron Microscopy: Methods and Protocols 2nd Edition (Methods in Molecular Biology Vol 369) PDF
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Additional resources for Electron Microscopy: Methods and Protocols 2nd Edition (Methods in Molecular Biology Vol 369)
Preparing Cultured Cells for TEM 13 Fig. 4. Streptococcal bacterial cells grown on the surface of a plastic Petri dish after culturing in liquid medium. The specimen was sectioned through a growing microcolony of bacteria. The bacteria exhibit different growth characteristics compared to growth in suspension culture as seen in Fig. 3. 1. Bar = 1 µm. 2. 1 M PIPES buffer, each for 5 min. 3. 2 M PIPES buffer stock. 4. Dehydrate in graded ethanol series, 10 min each. 14 Bozzola Fig. 5. Lab-Tek II plastic culture vessels consisting of Permanox slide surrounded by plastic chamber.
1. After 60 min fixation at room temperature, rinse the agar blocks in PIPES buffer for 10 min. 2. 1. following steps 8–14. 3. Pour epoxy 812 embedding medium into a Petri dish to a depth of several mm. If a flat embedding mold (of polypropylene or silicone) is available, use this instead of a Petri dish. 16 Bozzola Fig. 7. Portions of the separated epoxy layer containing embedded cells are broken into small pieces, or excised, and clamped into holder for ultramicrotomy. Arrow shows epoxy clamped into holder whereas arrowheads show areas of the epoxy that were excised using a cork borer.
Totowa, NJ 19 20 Mascorro and Bozzola infiltrated with a liquid resin, which is hardened by applying gentle heat (7,8). Hardened resin blocks containing the tissue specimens are thinly sectioned, and the sections treated with solutions of heavy metals in order to impart contrast (9,10). It is only at this point that the thin sections can be placed in the electron microscope column for observation and study. The purpose of chemical fixation is to stabilize the tissue so that it will present a final appearance not substantially unlike its original form.