Destructive tests on welds in metallic materials. Etchants for macroscopic and microscopic examination

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.

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SEM Examination of a Used Diamond Knife

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100066310 ◽

1971 ◽

Vol 29 ◽

pp. 452-453

Author(s):

J. Temple Black

Keyword(s):

Electron Microscope ◽

High Voltage ◽

High Magnification ◽

Metallic Materials ◽

Wide Spread ◽

Thin Sections ◽

Transmission Electron ◽

Scanning Transmission ◽

Scanning Electron ◽

Diamond Knife

Since its introduction by Fernandez-Moran, the diamond knife has gained wide spread usage as a common material for cutting of thin sections of biological and metallic materials into thin films for examination in the transmission electron microscope. With the development of high voltage E.M. and scanning transmission E.M., microtomy applications will become increasingly important in the preparation of specimens. For those who can afford it, the diamond knife will thus continue to be an important tool to accomplish this effort until a cheaper but equally strong and sharp tool is found to replace the diamond, glass not withstanding.In Figs. 1 thru 3, a first attempt was made to examine the edge of a used (β=45°) diamond knife by means of the scanning electron microscope. Because diamond is conductive, first examination was tried without any coating of the diamond. However, the contamination at the edge caused severe charging during imaging. Next, a thin layer of carbon was deposited but charging was still extensive at high magnification - high voltage settings. Finally, the knife was given a light coating of gold-palladium which eliminated the charging and allowed high magnification micrographs to be made with reasonable resolution.

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Artifact-free electron microscopic immunocytochemistry on rat brain tissue

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100085666 ◽

1991 ◽

Vol 49 ◽

pp. 272-273

Author(s):

Veronika Burmeister ◽

N. Ludvig ◽

P.C. Jobe

Keyword(s):

Microscopic Examination ◽

Free Electron ◽

Electron Microscopic Examination ◽

Ultrastructural Localization ◽

Rabbit Serum ◽

Electron Microscopic ◽

Electron Microscopic Immunocytochemistry ◽

Phosphate Buffered Saline ◽

Rat Brain Tissue ◽

Immune Rabbit

Electron microscopic immunocytochemistry provides an important tool to determine the ultrastructural distribution of various molecules in both normal and pathologic tissues. However, the specific immunostaining may be obscured by artifactual immunoreaction product, misleading the investigator. Previous observations show that shortening the incubation period with the primary antibody from the generally used 12-24 hours to 1 hour substantially reduces the artifactual immunostaining. We now extend this finding by the demonstration of artifact-free ultrastructural localization of the Ca2/calmodulindependent cyclic nucleotide phosphodiesterase (CaM-dependent PDE) immunoreactivity in brain.Anesthetized rats were perfused transcardially with phosphate-buffered saline followed by a fixative containing paraformaldehyde (4%) and glutaraldehyde (0.25%) in PBS. The brains were removed, and 40μm sections were cut with a vibratome. The sections were processed for immunocytochemistry as described by Ludvig et al. Both non-immune rabbit serum and specific CaM-dependent PDE antibodies were used. In both experiments incubations were at one hour and overnight. The immunostained sections were processed for electron microscopic examination.

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