By Robin, Ed. Harris
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Extra resources for Cell Biology Protocols
And Bracegirdle, B. (1998) Introduction to Light Microscopy, 2nd edn. Bios Scientific Publishers, Oxford. 2. Rubbi, C. ) (1994) Light Microscopy: Essential Data. John Wiley, London. 3. , Goldman, R. and Leinwand, L. (1998) Cells: A Laboratory Manual. Cold Spring Harbor Laboratory Press, New York. 4. Wright, S. and Wright, D. (2002) in Methods in Cell Biology, Vol. 70: Cell Biological Applications of Confocal Microscopy (B. ), pp. 1–85. Academic Press, New York. 5. Boon, M. and Drijver, J. (1986) Routine Cytological Staining Techniques: Theoretical Background and Practice.
5 Notes These procedures 30 min. 1 ); they are also available commercially. Quantofoil micromachined holey carbon grids can also be used. 2 These procedures, developed initially by Marc Adrian and his colleagues [7, 8] usually require a protein/lipid/ nucleic acid concentration somewhat in excess of 1 mg/ml. The study of specimens prepared by this technique requires a suitable cryoelectron microscope with cryoholder and cryotransfer system. Specimens have to be studied in the electron microscope under strict low electron dose conditions.
2). 13. Remove excess negative stain with a filter paper wedge and air-dry. This procedure takes approximately 2 h. 1 Attachment of cells to mica can be monitored by light microscopy. Brief 32 BASIC ELECTRON MICROSCOPY diam. g. 3 mm squares with the corners cut off). Also, droplet immunolabelling can be performed at this stage, either directly or after brief glutaraldehyde fixation as in. 1% glutaraldehyde in PBS. Avoid excessive fixation, as this will interfere with the cell cleavage. External labelling can be applied to mica-bound cells prior to cleavage.
Cell Biology Protocols by Robin, Ed. Harris