Critical Point Drying of Protozoan Cells and Other Biological Specimens for Scanning Electron Microscopy: Apparatus and Methods of Specimen Preparation

1974 ◽  
Vol 93 (1) ◽  
pp. 124 ◽  
Author(s):  
John J. Ruffolo
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Critical Point ◽  
Specimen Preparation ◽  
Critical Point Drying ◽  
Scanning Electron

Preservation of Plant Cell Surfaces For Scanning Electron Microscopy

1973 ◽  
Vol 31 ◽  
pp. 472-473
Author(s):  
Linda M. Sicko ◽  
Thomas E. Jensen
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Critical Point ◽  
Filter Paper ◽  
Freeze Drying ◽  
Cell Surfaces ◽  
Air Drying ◽  
Popular Method ◽  
Critical Point Drying ◽  
Scanning Electron

The use of critical point drying is rapidly becoming a popular method of preparing biological samples for scanning electron microscopy. The procedure is rapid, and produces consistent results with a variety of samples. The preservation of surface details is much greater than that of air drying, and the procedure is less complicated than that of freeze drying. This paper will present results comparing conventional air-drying of plant specimens to critical point drying, both of fixed and unfixed material. The preservation of delicate structures which are easily damaged in processing and the use of filter paper as a vehicle for drying will be discussed.

Intracellular structures of rapid frozen biological samples observed by high-resolution low-temperature Scanning Electron Microscopy

1989 ◽  
Vol 47 ◽  
pp. 736-737
Author(s):  
T. Inoué ◽  
H. Koike
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Low Temperature ◽  
Liquid Nitrogen ◽  
Specimen Preparation ◽  
Chemical Fixation ◽  
Brown Adipose ◽  
Critical Point Drying ◽  
Temperature Scanning ◽  
Scanning Electron

Low temperature scanning electron microscopy (LTSEM) is useful to avoid artifacts such as deformation and extraction, because specimens are not subjected to chemical fixation, dehydration and critical-point drying. Since Echlin et al developed a LTSEM, many techniques and instruments have been reported for observing frozen materials. However, intracellular structures such as mitochondria and endoplasmic reticulum have been unobservable by the method because of the low resolving power and inadequate specimen preparation methods. Recently, we developed a low temperature SEM that attained high resolutions. In this study, we introduce highly magnified images obtained by the newly developed LTSEM, especially intracellular structures which have been rapidly frozen without chemical fixation.[Specimen preparations] Mouse pancreas and brown adipose tissues (BAT) were used as materials. After the tissues were removed and cut into small pieces, the specimen was placed on a cryo-tip and rapidly frozen in liquid propane using a rapid freezing apparatus (Eiko Engineering Co. Ltd., Japan). After the tips were mounted on the specimen stage of a precooled cryo-holder, the surface of the specimen was manually fractured by a razor blade in liquid nitrogen. The cryo-holder was then inserted into the specimen chamber of the SEM (ISI DS-130), and specimens were observed at the accelerating voltages of 5-8 kV. At first the surface was slightly covered with frost, but intracellular structures were gradually revealed as the frost began to sublimate. Gold was then coated on the specimen surface while tilting the holder at 45-90°. The holder was connected to a liquid nitrogen reservoir by means of a copper braid to maintain low temperature.

Specimen chambers for critical point drying for scanning electron microscopy

1988 ◽  
Vol 8 (4) ◽  
pp. 443-444
Author(s):  
W. M. Hess ◽  
J. V. Allen ◽  
J. S. Gardner ◽  
Jay Reynolds
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Critical Point ◽  
Critical Point Drying ◽  
Scanning Electron

Comparison of preparation techniques of mixed samples (fungi–helminth eggs) for scanning electron microscopy by critical point drying

2006 ◽  
Vol 99 (4) ◽  
pp. 455-458 ◽  
Author(s):  
P. L. Sarmiento ◽  
María L. Ciarmela ◽  
P. Sánchez Thevenet ◽  
M. C. Minvielle ◽  
J. A. Basualdo
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Critical Point ◽  
Helminth Eggs ◽  
Critical Point Drying ◽  
Preparation Techniques ◽  
Mixed Samples ◽  
Scanning Electron

Delicate Botanical Specimens Preserved for Scanning Electron Microscopy by Critical Point Drying

1971 ◽  
Vol 29 ◽  
pp. 450-451
Author(s):  
Arthur L. Cohen ◽  
Gerald E. Garner
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Critical Point ◽  
Freeze Drying ◽  
Animal Cell ◽  
Pollen Grains ◽  
Scanning Electron Micrographs ◽  
Animal Cells ◽  
Critical Point Drying ◽  
Scanning Electron

The surface forms and structures of animal cells have been strikingly preserved for scanning electron microscopy by freeze-drying and by critical point drying both by the method with CO2 used as the transitional fluid and the later procedure which uses a fluorocarbon (Freon 13) as a medium for the transition from the liquid to the gaseous environment. Freeze-drying is often prolonged (5-12 hours as compared with an hour or less by the critical point method) and in our experience with mold cultures on agar, the substrate shrivels and cracks and hyphal filaments are distorted.Despite, and possibly because of a flexible but inelastic cell wall, plant cells often show greater distortion than do animal cells after evaporative drying or replacement dehydration for mixrotechnical work. The animal cell membrane can contract more or less uniformly on drying - as shown by the numerous micrographs of well-preserved erythrocytes, while plant cell walls often crumple. The many scanning electron micrographs of partially collapsed pollen grains bear witness to this fact.

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