By G. L. Nicolson (auth.), James K. Koehler Ph. D. (eds.)
The use of the time period "advanced" within the identify of this booklet is a bit ar bitrary and intensely a lot relative with admire to time. Many options that have been thought of on the "cutting side" of ultrastructural technique quite a few years in the past are actually rou tin ely utilized in a variety of laboratories. you may cite freeze-fracture, cryothin sectioning, or certainly lots of the box of test ning electron microscopy as concrete examples. therefore using the time period "ad vanced recommendations" needs to be interpreted in regards to the current cutting-edge, and comes in handy in simple terms in informing the capability reader that this quantity isn't really a primer for use as an preliminary advent into easy organic elec tron microscopy. many fantastic volumes have crammed that area of interest long ago few years, and it isn't meant that this modest e-book be an entire com pendium of the sphere. moreover, any constrained collection of papers on advanc ed ideas unavoidably displays the personal tastes and arbitrary whims of the editor, thereby aside from many both vital methods which the an expert reader will conveniently establish. the 1st quantity of this sequence seemed nearly 5 years in the past and illustrated suggestions which have been notion to symbolize complicated and but ba sically morphological equipment for gaining elevated ultrastructural informa tion from organic specimens. the current quantity, nevertheless, stresses thoughts which offer particular physicochemical facts at the speci mens as well as the structural information.
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Extra info for Advanced Techniques in Biological Electron Microscopy II: Specific Ultrastructural Probes
Cell BioI. (submitted, 1977) Nicolson, G. : Mobility and the restriction of mobility of plasma membrane lectin-binding components. Science 184, 1294 -1296 (1974) Nicolson, G. ]. Cell BioI. : Lymphocyte transformation induced by concanavalin A and its reversion by methyl-alpha-D-mannopyranoside. Biochim. Biophys. , Barondes, S. : Agglutinin from Limulus polyphemus: purification with formalinized horse erythrocytes as the affinity adsorbent. Biochim. Biophys. Acta 393, 115 - 123 (1975) Oikawa, T, Nicolson, G.
E) Control SV 40-transformed hamster cell labeled with Con A, then peroxidase at 22° C, and immediately fixed. (f, g) SV 40-transformed hamster cells labeled at 22° C and postincubated at 37° C for 15 and 60 min, respectively. (From HUET and BERNHARD, 1974, courtesy of Dr. C. HUET) X 14,000 24 ·G. L. NICOLSON postincubated at 37° C, lectin-induced redistribution occurred (Fig. 5 b, c), eventually leading to endocytosis of lectin-peroxidase complexes by 60 min (Fig. 5 d). When HUETand BERNHARD (1974) examined an SV 40-transformed hamster embryo cell line (Cl 2TSVs), these events occurred even more rapidly than on the untransformed cell surface (Fig.
The fixed samples are treated for 15 to 30 min in the dark with a solution of 3,3'-diaminobenzidine hydrochloride (Sigma Chemical, St. 4 containing 1 drop of 30% HP2 per 5 mlof Table 5. Some uses of lectin-peroxidase techniques Lectin(s) Biological material Con A Burkitt tumor and intestinal mucosa AVRAMEAS (1970); BERNHARD and AVRAMEAS (1971) Fibroblast movement ABERCROMBIE et al. (1972) Normal and polyoma-transformed MARTINEZ-PALOMO et al. cells (1972) Serum components adherent to ROWLATT and WICKER (1972) substrate Normal and SV40-transformed cells BRETTON et al.