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Cell Biology Protocols by Robin, Ed. Harris PDF

By Robin, Ed. Harris

ISBN-10: 0470847581

ISBN-13: 9780470847589

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Extra resources for Cell Biology Protocols

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E. without primary/secondary antibodies, should be incorporated alongside grids passed through the complete protocol. 11 Imaging the nuclear matrix and cytoskeleton by embedment-free electron microscopy Jeffrey A. Nickerson and Jean Underwood Introduction Conventional embedded section electron microscopy is well suited for the visualization in cells and tissues of membranebounded organelles, whose sectioning generates characteristic membrane profiles. Among other uses, embedded sections also allow simple selective staining for cell components using electron dense elements, such as EDTA-regressive or Terbium staining for RNA [16, 17] and allow the imaging of chromatin packaging densities, for example, distinguishing euchromatin and heterochromatin.

R. ) (1991) Electron Microscopy in Biology: A Practical Approach. IRL Press at Oxford University Press, Oxford. 2. Sommerville, J. and U. Scheer (eds) (1987) Electron Microscopy in Molecular Biology: A Practical Approach, IRL Press, Oxford. 3. Harris, J. R. (1997) Negative Staining and Cryoelectron Microscopy: The Thin Film Techniques. RMS Handbook No. 35, BIOS Scientific Publishers, Oxford. 4. Hajibagheri, M. A. N. ) (1999) Electron Microscopy Methods and Protocols. Humana Press, Totowa, New Jersey, USA.

To stain sections with uranyl acetate 2 : put 20 µl droplets of uranyl acetate solution onto paraffin wax or Parafilm in a Petri dish. 2. Float the grids with section side down 3 on the stain droplets in a covered Petri dish. The usual staining time ranges from 30 min to 2 h at room temperature. 3. Wash the grids thoroughly individually with flowing distilled water from a pipette, while holding them in forceps, or rinse in three consecutive 100 ml beakers of distilled water. 4. Stain sections with lead citrate: place some NaOH pellets (to minimize atmospheric CO2 ) in the Petri dish alongside 40 µl droplets of freshly filtered/ centrifuged stain solution.

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Cell Biology Protocols by Robin, Ed. Harris


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