Innovative Technologies in Advanced SIMS : Toward Organic and Biological Material Analysis

Color contrasting of 1 to 2 micron sections of plastic embedded biological material is an important adjunct to electron microscopy. The procedures in general use today are simple and rapid giving monochromatic results, e.g., toluidine blue. Although many di- and polychromatic histologic staining techniques have been modified to obtain a counterstaining effect with plasticembedded tissue, the methods are usually undesirable for routine work because they are time consuming, complicated and often defy good reproducibility.

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Ultrathin frozen sections of yeast without aldehyde prefixation

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100081152 ◽

1978 ◽

Vol 36 (2) ◽

pp. 58-59

Author(s):

K. J. Böhm ◽

a. E. Unger

Keyword(s):

Aqueous Solutions ◽

Biological Material ◽

Yeast Cells ◽

Frozen Sections ◽

Good Preservation ◽

Ultrathin Frozen Sections

During the last years it was shown that also by means of cryo-ultra-microtomy a good preservation of substructural details of biological material was possible. However the specimen generally was prefixed in these cases with aldehydes.Preparing ultrathin frozen sections of chemically non-prefixed material commonly was linked up to considerable technical and manual expense and the results were not always satisfying. Furthermore, it seems to be impossible to carry out cytochemical investigations by means of treating sections of unfixed biological material with aqueous solutions.We therefore tried to overcome these difficulties by preparing yeast cells (S. cerevisiae) in the following manner:

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Albumins as Embedding Media for Electron Microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100064025 ◽

1969 ◽

Vol 27 ◽

pp. 422-423 ◽

Cited By ~ 1

Author(s):

J. L. Farrant ◽

J. D. McLean

Keyword(s):

Electron Microscopy ◽

Electron Microscope ◽

High Temperature ◽

Biological Material ◽

Globular Proteins ◽

The Past ◽

Antibody Staining ◽

Thin Sectioning ◽

Embedding Medium

For electron microscope techniques such as ferritin-labeled antibody staining it would be advantageous to have available a simple means of thin sectioning biological material without subjecting it to lipid solvents, impregnation with plastic monomers and their subsequent polymerization. With this aim in view we have re-examined the use of protein as an embedding medium. Gelatin which has been used in the past is not very satisfactory both because of its fibrous nature and the high temperature necessary to keep its solutions fluid. We have found that globular proteins such as the serum and egg albumins can be cross-linked so as to yield blocks which are suitable for ultrathin sectioning.

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Ferritin Labeled Antibody Staining of Thin Sections

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100064013 ◽

1969 ◽

Vol 27 ◽

pp. 420-421

Author(s):

J. D. McLean ◽

S. J. Singer

Keyword(s):

Biological Material ◽

Water Soluble ◽

Thin Sections ◽

Antibody Staining ◽

Thin Sectioning ◽

Ultrathin Sections

The successful application of ferritin labeled antibodies (F-A) to ultrathin sections of biological material has been hampered by two main difficulties. Firstly the normally used procedures for the preparation of material for thin sectioning often result in a loss of antigenicity. Secondly the polymers employed for embedding may non-specifically absorb the F-A. Our earlier use of cross-linked polyampholytes as embedding media partially overcame these problems. However the water-soluble monomers used for this method still extract many lipids from the material.

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Material analysis techniques using the ion mill

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100167639 ◽

1996 ◽

Vol 54 ◽

pp. 1034-1035

Author(s):

J. P. Benedict ◽

R. M. Anderson ◽

S. J. Klepeis

Keyword(s):

Polished Surface ◽

Surface Material ◽

Analysis Techniques ◽

Material Analysis ◽

Optical Inspection ◽

Electron Microscopy Analysis ◽

Microscopy Analysis ◽

Argon Ions ◽

The Impact ◽

Scanning Electron

Ion mills equipped with flood guns can perform two important functions in material analysis; they can either remove material or deposit material. The ion mill holder shown in Fig. 1 is used to remove material from the polished surface of a sample for further optical inspection or SEM ( Scanning Electron Microscopy ) analysis. The sample is attached to a pohshing stud type SEM mount and placed in the ion mill holder with the polished surface of the sample pointing straight up, as shown in Fig 2. As the holder is rotating in the ion mill, Argon ions from the flood gun are directed down at the top of the sample. The impact of Argon ions against the surface of the sample causes some of the surface material to leave the sample at a material dependent, nonuniform rate. As a result, the polished surface will begin to develop topography during milling as fast sputtering materials leave behind depressions in the polished surface.

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