Processing Techniques for Scanning Electron Microscopy Imaging of Giant Cells from Giant Cell Tumors of Bone

2019 ◽  
Vol 25 (6) ◽  
pp. 1376-1382
Author(s):  
Asit Ranjan Mridha ◽  
Indu Barwal ◽  
Abhishek Gupta ◽  
Abdul Majeed ◽  
Adarsh W. Barwad ◽  
...  
Keyword(s):  
Surface Morphology ◽  
Critical Point ◽  
Giant Cell ◽  
Giant Cells ◽  
Giant Cell Tumors ◽  
Microscopy Imaging ◽  
Air Drying ◽  
Critical Point Drying ◽  
Sem Imaging ◽  
Scanning Electron

AbstractGiant cell tumor (GCT) of bone is a common benign lesion that causes significant morbidity due to the failure of modern medical and surgical treatment. Surface ultra-structures of giant cells (GCs) may help in distinguishing aggressive tumors from indolent GC lesions. This study aimed to standardize scanning electron microscopic (SEM) imaging of GC from GCT of bone. Fresh GCT collected in Dulbecco's Modified Eagle Medium was washed to remove blood, homogenized, or treated with collagenase to isolate the GCs. Mechanically homogenized and collagenase-digested GCs were imaged on SEM after commonly used drying methodologies such as air-drying, tetramethylsilane (TMS)-drying, freeze-drying, and critical point-drying (CPD) for the optimization of sample processing. The collagenase-treated samples yielded a greater number of isolated GC and showed better surface morphology in comparison to mechanical homogenization. Air-drying was associated with marked cell shrinkage, and freeze-dried samples showed severe cell damage. TMS methodology partially preserved the cell contour and surface structures, although the cell shape was distorted. GC images with optimum surface morphology including membrane folding and microvesicular structures on the surface were observed only in collagenase-treated and critical point-dried samples. Collagenase digestion and critical point/TMS-drying should be performed for optimal SEM imaging of individual GCs.

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.

Biological Sample Preparation for SEM Imaging of Porcine Retina

2012 ◽  
Vol 20 (2) ◽  
pp. 28-31 ◽  
Author(s):  
Patrick Moran ◽  
Brittany Coats
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Sample Preparation ◽  
Critical Point ◽  
Biological Sample ◽  
Environmental Scanning Electron Microscopy ◽  
Preparation Methods ◽  
Critical Point Drying ◽  
Sem Imaging ◽  
Scanning Electron

Sample preparation is a critical step in scanning electron microscopy (SEM) imaging. This is especially true for biological samples because of charge build-up and sensitivity to vacuum and electron beam damage. In terms of ultrastructure imaging, a variety of advancements in detectors and approaches have improved biological imaging such that fewer steps are required for sample preparation. However, the conventional approach incorporating osmium tetroxide fixing, ethanol dehydrating, critical-point drying, and coating still finds useful application. This paper evaluates three biological sample-preparation methodologies for imaging the ultrastructure of immature porcine retina. The three preparation methods examined are critical-point drying (CPD), hexamethyldisilazane (HMDS) dehydration, and direct imaging by environmental scanning electron microscopy (ESEM). Preparation methodologies were evaluated based on resulting image quality and reduced potential for artifacts.

The Critical Point Drying Method Applied to Scanning Electron Microscopy of L-929 Cells

1970 ◽  
Vol 28 ◽  
pp. 278-279
Author(s):  
Charles TurnbiLL ◽  
Delbert E. Philpott
Keyword(s):  
Electron Microscopy ◽  
Carbon Dioxide ◽  
Surface Tension ◽  
Critical Point ◽  
Freeze Drying ◽  
Attractive Alternative ◽  
Crystal Formation ◽  
Critical Point Drying ◽  
Time Required ◽  
Scanning Electron

The advent of the scanning electron microscope (SCEM) has renewed interest in preparing specimens by avoiding the forces of surface tension. The present method of freeze drying by Boyde and Barger (1969) and Small and Marszalek (1969) does prevent surface tension but ice crystal formation and time required for pumping out the specimen to dryness has discouraged us. We believe an attractive alternative to freeze drying is the critical point method originated by Anderson (1951; for electron microscopy. He avoided surface tension effects during drying by first exchanging the specimen water with alcohol, amy L acetate and then with carbon dioxide. He then selected a specific temperature (36.5°C) and pressure (72 Atm.) at which carbon dioxide would pass from the liquid to the gaseous phase without the effect of surface tension This combination of temperature and, pressure is known as the "critical point" of the Liquid.

The Effects of Drying Techniques on Clay-Rich Soil Texture

1974 ◽  
Vol 32 ◽  
pp. 466-467
Author(s):  
T. G. Naymik
Keyword(s):  
Critical Point ◽  
Liquid Nitrogen ◽  
Liquid Nitrogen Temperature ◽  
Drying Methods ◽  
Air Drying ◽  
Freeze Dried ◽  
Critical Point Drying ◽  
Set Up ◽  
The Relationship ◽  
Quartz Grains

Three techniques were incorporated for drying clay-rich specimens: air-drying, freeze-drying and critical point drying. In air-drying, the specimens were set out for several days to dry or were placed in an oven (80°F) for several hours. The freeze-dried specimens were frozen by immersion in liquid nitrogen or in isopentane at near liquid nitrogen temperature and then were immediately placed in the freeze-dry vacuum chamber. The critical point specimens were molded in agar immediately after sampling. When the agar had set up the dehydration series, water-alcohol-amyl acetate-CO2 was carried out. The objectives were to compare the fabric plasmas (clays and precipitates), fabricskeletons (quartz grains) and the relationship between them for each drying technique. The three drying methods are not only applicable to the study of treated soils, but can be incorporated into all SEM clay soil studies.

Rapid Critical Point Drying of Tissues in a Parr Bomb

1972 ◽  
Vol 30 ◽  
pp. 400-401
Author(s):  
C.A. Baechler ◽  
W. C. Pitchford ◽  
J. M. Riddle ◽  
C.B. Boyd ◽  
H. Kanagawa ◽  
...  
Keyword(s):  
Critical Point ◽  
Critical Pressure ◽  
Soft Tissues ◽  
Biological Tissues ◽  
Critical Temperatures ◽  
Air Drying ◽  
The Past ◽  
Critical Point Drying ◽  
Great Stress ◽  
Infiltration Techniques

Preservation of the topographic ultrastructure of soft biological tissues for examination by scanning electron microscopy has been accomplished in the past by using lengthy epoxy infiltration techniques, or dehydration in ethanol or acetone followed by air drying. Since the former technique requires several days of preparation and the latter technique subjects the tissues to great stress during the phase change encountered during air-drying, an alternate rapid, economical, and reliable method of surface structure preservation was developed. Turnbill and Philpott had used a fluorocarbon for the critical point drying of soft tissues and indicated the advantages of working with fluids having both moderately low critical pressures as well as low critical temperatures. Freon-116 (duPont) which has a critical temperature of 19. 7 C and a critical pressure of 432 psi was used in this study.

Pulmonary microcirculation: tubules rather than sheet and post

1982 ◽  
Vol 53 (2) ◽  
pp. 510-515 ◽  
Author(s):  
W. G. Guntheroth ◽  
D. L. Luchtel ◽  
I. Kawabori
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Critical Point ◽  
Ringer Solution ◽  
Basic Component ◽  
Pulmonary Vasculature ◽  
Critical Point Drying ◽  
Pulmonary Microcirculation ◽  
Scanning Electron

We examined latex casts of the pulmonary microcirculation with the scanning electron microscope (SEM). Mature rats were anesthetized and ventilated; the pulmonary vasculature was washed out with lactated Ringer solution and then filled with a mixture of Geon latexes. The airways were filled with glutaraldehyde with resulting transmural vascular pressures of 10 cmH2O. After critical-point drying and corrosive removal of the lung tissue, SEM studies of the vascular replicas revealed two distinct patterns of pulmonary microcirculation: 1) sparse, long, tubular capillaries that comprise the thin subpleural layer and appear as “filler” in the peribronchial spaces; and 2) alveolar microcirculation that is composed of tightly matted, intersecting tubules, shorter but of the same diameter as type 1, in spherical array in two layers. The alveolar capillaries at low magnification appear superficially as sheets; however, the detailed morphology is not consistent with the sheet-and-post model. We conclude that the basic component of the pulmonary microcirculation is tubular and not different from other capillary beds except in density.

A comparison of microstructures of Cheddar cheese curd manufactured with calf rennet, bovine pepsin, and porcine pepsin

1976 ◽  
Vol 43 (1) ◽  
pp. 113-115 ◽  
Author(s):  
M. F. Eino ◽  
D. A. Biggs ◽  
D. M. Irvine ◽  
D. W. Stanley
Keyword(s):  
Electron Microscope ◽  
Scanning Electron Microscope ◽  
Critical Point ◽  
Cheddar Cheese ◽  
Microscope Examination ◽  
Porcine Pepsin ◽  
Electron Microscope Examination ◽  
Critical Point Drying ◽  
Cheese Curd ◽  
Scanning Electron

SummaryCalf rennet, bovine pepsin, and porcine pepsin were used to produce cheese curd, using the same milk and lactic culture for each. Specimens were prepared for scanning electron microscope examination by a modified critical-point drying technique.From examination of the micrographs, the curd made with bovine and porcine pepsin were similar in structure and in orientation of the coagulated protein, whereas the curd produced with rennet was different, having a more compact and organized structure.

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