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7 mL glass processing bottles with lids. 6 Epoxy Resins (Epon) 1. , Agar 100 resin, TAAB resin. HAZARD 2. DDSAdodecenyl succinic anhydride. HAZARD 3. MNAmethyl nadic anhydride. HAZARD 4. BDMAbenzyl dimethylamine. HAZARD. 5. Low density polyethylene bottles 50 mL. 7 Dehydration Solvents Use Analar reagents. 1. 30, 50,70, 90% ethanol or acetone in distilled water. 2. 100% ethanol or acetone. 3. 100% ethanol or acetone with added molecular sieve 5a. Do not disturb the molecular sieve. 8 Infiltration and Embedding 1.

R. and Hobot, J. A. (1993) Resin Microscopy and On-Section Immunocytochemistry, Springer-Veriag, Berlin, Germany. Page 13 2 Negative Staining of Thinly Spread Biological Particulates J. Robin Harris 1 Introduction The negative staining of virus particles for TEM study was introduced in the late 1950s, following the establishment of a standardized procedure by Brenner and Horne in 1959 (1). , nonaggregated) material was available. In addition to viruses, this material ranged from purified enzymes and other soluble protein molecules and components of molecular mass in the range of 200 kDa up to several MDa (such as the molluscan hemocyanins and ribosomes), to isolated cellular organelles, membrane fractions, bacterial cell walls and membranes, and filamenous protein structures of many types, also liposomal and reconstituted membrane systems, and even synthetic polymers.

In addition, the combination of the established techniques of cryoelectron microscopy for the study of unstained biological materials with those of negative staining has opened up new and exiting possibilities (4; see also Chapter 3). It is the aim of this Chapter to present some of the well-established and newer procedures (5,6) for air-dry negative staining on continuous thin carbon support films and across small holes in carbon films, with indication of the varying possibilities, applications and technical limitations.

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