Isolation of high-rate DNA synthetic cells by Con A chromatography

Blast cells which were derived from rat lymphocytes by stimulation with phytohemagglutinin (PHA), concanavalin A (Con A), or pokeweed mitogen (PWM) transformed within 2–3 days into a new type of lymphocytes when plated without mitogen on embryo fibroblast monolayers. These lymphocytes were termed secondary lyrophocytes. Upon addition of PWM to PWM-secondary lymphocytes a marked adherence to fibroblast monolayers was observed. The degree of adherence was estimated (a) by direct count of the lymphocytes in the medium and in the trypsinized fibroblast fraction, and (b) by using 51Cr-labeled lymphocytes. The adherence process required incubation at 37°C. The process started immediately after the addition of PWM and reached a plateau at 6 hr. At this time more than 80% of the lymphocytes adhered. In the absence of PWM only 12% of the lymphocytes were found in the fibroblast fraction. Unlike PWM-lymphocytes. Con A-lymphocytes, PHA-lymphocytes, and ordinary lymphocytes taken directly from the rat lymph nodes adhered only slightly more in the presence of PWM (10–20% adherence of ordinary lymphocytes) than in its absence (8% adherence). The adherence of the secondary lymphocytes and the ordinary lymphocytes was also studied in the presence of Con A and PHA. These mitogens induced high rate of adherence and they did not demonstrate specificity in their action. The adherence was accompanied by transformation of the lymphocytes to blast cells endowed with target-cell lytic ability. This transformation occurred mostly in the adhering fraction of the lymphocyte population. The results support the notion that target-cell recognition and destruction in cellular immunity involve contact between the cells.

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Structure of Palladium Single-Crystal Films Prepared by Flash Evaporation onto (001) NaCl Substrates

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100069272 ◽

1970 ◽

Vol 28 ◽

pp. 456-457

Author(s):

L. E. Murr ◽

G. Wong

Keyword(s):

Single Crystal ◽

High Vacuum ◽

Ultra High Vacuum ◽

High Rate ◽

Flash Evaporation ◽

Optimum Load ◽

Single Crystal Films ◽

Crystal Films ◽

A Current

Palladium single-crystal films have been prepared by Matthews in ultra-high vacuum by evaporation onto (001) NaCl substrates cleaved in-situ, and maintained at ∼ 350° C. Murr has also produced large-grained and single-crystal Pd films by high-rate evaporation onto (001) NaCl air-cleaved substrates at 350°C. In the present work, very large (∼ 3cm2), continuous single-crystal films of Pd have been prepared by flash evaporation onto air-cleaved (001) NaCl substrates at temperatures at or below 250°C. Evaporation rates estimated to be ≧ 2000 Å/sec, were obtained by effectively short-circuiting 1 mil tungsten evaporation boats in a self-regulating system which maintained an optimum load current of approximately 90 amperes; corresponding to a current density through the boat of ∼ 4 × 104 amperes/cm2.

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Lectin Binding Studies on RNA Tumor Virus-Infected Cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100115250 ◽

1975 ◽

Vol 33 ◽

pp. 326-327

Author(s):

D. C. Hixson

Keyword(s):

Binding Sites ◽

Lectin Binding ◽

Culture Conditions ◽

3T3 Cells ◽

Binding Studies ◽

Con A ◽

Microscopic Studies ◽

Tumor Virus ◽

Infected Cells ◽

Cell Culture Conditions

The abilities of plant lectins to preferentially agglutinate malignant cells and to bind to specific monosaccharide or oligosaccharide sequences of glycoproteins and glycolipids make them a new and important biochemical probe for investigating alterations in plasma membrane structure which may result from malignant transformation. Electron and light microscopic studies have demonstrated clustered binding sites on surfaces of SV40-infected or tryp- sinized 3T3 cells when labeled with concanavalin A (con A). No clustering of con A binding sites was observed in normal 3T3 cells. It has been proposed that topological rearrangement of lectin binding sites into clusters enables con A to agglutinate SV40-infected or trypsinized 3T3 cells (1). However, observations by other investigators have not been consistent with this proposal (2) perhaps due to differences in reagents used, cell culture conditions, or labeling techniques. The present work was undertaken to study the lectin binding properties of normal and RNA tumor virus-infected cells and their associated viruses using lectins and ferritin-conjugated lectins of five different specificities.

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Rapid Freezing of Freeze-Etch Specimens

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s1431927600003275 ◽

1980 ◽

Vol 38 ◽

pp. 752-755

Author(s):

A. Elgsaeter ◽

T. Espevik ◽

G. Kopstad

Keyword(s):

Low Temperature ◽

Metal Surface ◽

Relative Velocity ◽

Thermal Contact ◽

High Rate ◽

Rapid Freezing ◽

Temperature Decrease ◽

Freeze Etching

The importance of a high rate of temperature decrease (“rapid freezing”) when freezing specimens for freeze-etching has long been recognized1. The two basic methods for achieving rapid freezing are: 1) dropping the specimen onto a metal surface at low temperature, 2) bringing the specimen instantaneously into thermal contact with a liquid at low temperature and subsequently maintaining a high relative velocity between the liquid and the specimen. Over the last couple of years the first method has received strong renewed interest, particularily as the result of a series of important studies by Heuser and coworkers 2,3. In this paper we will compare these two freezing methods theoretically and experimentally.

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