Immuno-Fluorescence Scanning Electron Microscopy of Biological Cells

2010 ◽  
Vol 18 (5) ◽  
pp. 8-13 ◽  
Author(s):  
Phyllis J. Fisher ◽  
William S. Wessels ◽  
Allan B. Dietz ◽  
Franklyn G. Prendergast
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Function ◽  
Three Dimensional ◽  
Biological Cells ◽  
Cell Surfaces ◽  
Interaction Range ◽  
Spatial Control ◽  
Scanning Electron ◽  
Surface Changes

Scanning electron microscopy (SEM) can produce striking three-dimensional images of biological cells and tissues with submicron resolution of surface morphology. Such cell surfaces are often complex blends of folds, extrusions, and pockets that may be necessary in the positioning of specific molecules within interaction range of each other. Thus, surface changes can have a spatial control over some molecular functions, and identification of select molecules at distinct morphological locations becomes critical to our understanding of total cell function.

Biodegradable Cell Transplantation Devices for Tissue Regeneration

1991 ◽  
Vol 252 ◽  
Author(s):  
Antonios G. Mikos ◽  
Heidi L. Wald ◽  
Georgios Sarakinos ◽  
Susan M. Leite ◽  
Robert Langer
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Transplantation ◽  
Tissue Regeneration ◽  
Cell Function ◽  
Mercury Porosimetry ◽  
Three Dimensional ◽  
Visual Evaluation ◽  
Highly Porous ◽  
Scanning Electron

ABSTRACTBiodegradable polymers can be utilized as templates for cell transplantation and regeneration of metabolic organs and structural tissues. Candidate materials must be adhesive substrates for cells, promote cell growth and allow for retention of cell function. However, the processing requirements of such materials into highly porous three-dimensional structures with large surface per volume and an interconnecting pore network limits their potential application for tissue regeneration. A new processing technique was developed to produce uniform, three-dimensional cell transplantation devices of poly(lactic-co-glycolic acid). The process involved the preparation of highly porous membranes by a solvent-casting and particulate-leaching technique followed by their lamination. The device structural and mechanical properties depended on those of their constituent membranes, as evaluated by mercury porosimetry, scanning electron microscopy, and thermomechanical analysis. Cells to be seeded into the devices were injected from catheters incorporated within their structure. In vitro studies with model suspensions of dyed microspheres allowed for visual evaluation of the internal pore structure of various layered devices. From these studies, numerous parameters of device design for cell seeding were determined including pore size and injection rate. The membrane lamination technique produced devices without interfaces between layers as determined by microsphere injection and scanning electron microscopy.

Scanning and Transmission Microscopy of Nematocyst Batteries in Epitheliomuscular Cells of Hydra

1971 ◽  
Vol 29 ◽  
pp. 410-411 ◽  
Author(s):  
Jane A. Westfall ◽  
S. Yamataka ◽  
Paul D. Enos
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Osmium Tetroxide ◽  
External Surface ◽  
Three Dimensional ◽  
Surface Structures ◽  
Transmission Microscopy ◽  
Transmission Electron ◽  
Freeze Dried ◽  
Scanning Electron

Scanning electron microscopy (SEM) provides three dimensional details of external surface structures and supplements ultrastructural information provided by transmission electron microscopy (TEM). Animals composed of watery jellylike tissues such as hydras and other coelenterates have not been considered suitable for SEM studies because of the difficulty in preserving such organisms in a normal state. This study demonstrates 1) the successful use of SEM on such tissue, and 2) the unique arrangement of batteries of nematocysts within large epitheliomuscular cells on tentacles of Hydra littoralis.Whole specimens of Hydra were prepared for SEM (Figs. 1 and 2) by the fix, freeze-dry, coat technique of Small and Màrszalek. The specimens were fixed in osmium tetroxide and mercuric chloride, freeze-dried in vacuo on a prechilled 1 Kg brass block, and coated with gold-palladium. Tissues for TEM (Figs. 3 and 4) were fixed in glutaraldehyde followed by osmium tetroxide. Scanning micrographs were taken on a Cambridge Stereoscan Mark II A microscope at 10 KV and transmission micrographs were taken on an RCA EMU 3G microscope (Fig. 3) or on a Hitachi HU 11B microscope (Fig. 4).

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.

Scanning Electron Microscopy and Electron Microprobe Analysis of Malignant Cell Cultures

1968 ◽  
Vol 26 ◽  
pp. 164-165
Author(s):  
R. I. Johnsson-Hegyeli ◽  
A. F. Hegyeli ◽  
D. K. Landstrom ◽  
W. C. Lane
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Cell Cultures ◽  
Electron Microprobe ◽  
Microprobe Analysis ◽  
Electron Microprobe Analysis ◽  
Three Dimensional ◽  
Malignant Cell ◽  
Interference Microscopy ◽  
Scanning Electron

Last year we reported on the use of reflected light interference microscopy (RLIM) for the direct color photography of the surfaces of living normal and malignant cell cultures without the use of replicas, fixatives, or stains. The surface topography of living cells was found to follow underlying cellular structures such as nuceloli, nuclear membranes, and cytoplasmic organelles, making possible the study of their three-dimensional relationships in time. The technique makes possible the direct examination of cells grown on opaque as well as transparent surfaces. The successful in situ electron microprobe analysis of the elemental composition and distribution within single tissue culture cells was also reported.This paper deals with the parallel and combined use of scanning electron microscopy (SEM) and the two previous techniques in a study of living and fixed cancer cells. All three studies can be carried out consecutively on the same experimental specimens without disturbing the cells or their structural relationships to each other and the surface on which they are grown. KB carcinoma cells were grown on glass coverslips in closed Leighto tubes as previously described. The cultures were photographed alive by means of RLIM, then fixed with a fixative modified from Sabatini, et al (1963).

Forensic Identifications Aided by Scanning Electron Microscopy of Silicone-Epoxy Microreplicas of Calcified and Cornified Structures

1975 ◽  
Vol 33 ◽  
pp. 678-679 ◽  
Author(s):  
R.F. Sognnaes
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Forensic Science ◽  
George Washington ◽  
Epithelial Surface ◽  
Extended Application ◽  
Tissue Surfaces ◽  
Non Destructive ◽  
Scanning Electron ◽  
Surface Changes

Sufficient experience has been gained during the past five years to suggest an extended application of microreplication and scanning electron microscopy to problems of forensic science. The author's research was originally initiated with a view to develop a non-destructive method for identification of materials that went into objects of art, notably ivory and ivories. This was followed by a very specific application to the identification and duplication of the kinds of materials from animal teeth and tusks which two centuries ago went into the fabrication of the ivory dentures of George Washington. Subsequently it became apparent that a similar method of microreplication and SEM examination offered promise for a whole series of problems pertinent to art, technology and science. Furthermore, what began primarily as an application to solid substances has turned out to be similarly applicable to soft tissue surfaces such as mucous membranes and skin, even in cases of acute, chronic and precancerous epithelial surface changes, and to post-mortem identification of specific structures pertinent to forensic science.

scholarly journals Three-dimensional relationships between tumor cells and microcirculation with double cyanine immunolabeling, laser scanning confocal microscopy, and computer-assisted reconstruction: an alternative to cast corrosion preparations.

1994 ◽  
Vol 42 (5) ◽  
pp. 681-686 ◽  
Author(s):  
V Rummelt ◽  
L M Gardner ◽  
R Folberg ◽  
S Beck ◽  
B Knosp ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Scanning Electron Microscopy ◽  
Blood Vessels ◽  
Tumor Cells ◽  
Laser Scanning ◽  
Three Dimensional ◽  
Melanoma Cells ◽  
Tumor Blood Vessels ◽  
The Relationship ◽  
Scanning Electron

The morphology of the microcirculation of uveal melanomas is a reliable market of tumor progression. Scanning electron microscopy of cast corrosion preparations can generate three-dimensional views of these vascular patterns, but this technique sacrifices the tumor parenchyma. Formalin-fixed wet tissue sections 100-150 microns thick from uveal melanomas were stained with the lectin Ulex europaeus agglutinin I (UEAI) and proliferating cell nuclear antigen (PCNA) to demonstrate simultaneously the tumor blood vessels and proliferating tumor cells. Indocarbocyanine (Cy3) was used as a fluorophore for UEAI and indodicarbocyanine (Cy5) was used for PCNA. Double labeled sections were examined with a laser scanning confocal microscope. Images of both stains were digitized at the same 5-microns intervals and each of the two images per interval was combined digitally to form one image. These combined images were visualized through voxel processing to study the relationship between melanoma cells expressing PCNA and various microcirculatory patterns. This technique produces images comparable to scanning electron microscopy of cast corrosion preparations while permitting simultaneous localization of melanoma cells expressing PCNA. The microcirculatory tree can be viewed from any perspective and the relationship between tumor cells and the tumor blood vessels can be studied concurrently in three dimensions. This technique is an alternative to cast corrosion preparations.

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