Albumins as Embedding Media for Electron Microscopy

1969 ◽  
Vol 27 ◽  
pp. 422-423 ◽  
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.

Ferritin Labeled Antibody Staining of Thin Sections

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.

scholarly journals ELECTRON MICROSCOPY OF THE NUCLEAR MEMBRANE OF AMOEBA PROTEUS

1956 ◽  
Vol 2 (4) ◽  
pp. 445-448 ◽  
Author(s):  
Marie H. Greider ◽  
Wencel J. Kostir ◽  
Walter J. Frajola
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Nuclear Membrane ◽  
Outer Layer ◽  
Microscope Study ◽  
Electron Microscope Study ◽  
Cell Types ◽  
Amoeba Proteus ◽  
Nuclear Membranes ◽  
Thin Sectioning

An electron microscope study of the nuclear membrane of Amoeba proteus by thin sectioning techniques has revealed an ultrastructure in the outer layer of the membrane that is homologous to the pores and annuli observed in the nuclear membranes of many other cell types studied by these techniques. An inner honeycombed layer apparently unique to Amoeba proteus is also described.

scholarly journals THE USE OF SPECIFIC ANTIBODY IN ELECTRON MICROSCOPY

1961 ◽  
Vol 11 (3) ◽  
pp. 533-547 ◽  
Author(s):  
Frank A. Pepe ◽  
H. Finck ◽  
H. Holtzer
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Fluorescence Microscopy ◽  
Adenosine Triphosphate ◽  
Potassium Iodide ◽  
Specific Antibody ◽  
Breast Muscle ◽  
Chicken Breast ◽  
Antibody Staining

Antibody staining was observed in the electron microscope by means of untagged antibody and osmium fixation. The antibody was visualized as a change in morphology due to its deposition on the antigenic structures. Glycerinated chicken breast muscle was stained with antimyosin, anti-H-meromyosin, and antiactin. The staining patterns obtained by electron microscopy were consistent with those previously demonstrated by fluorescence microscopy. A second method was used for confirmation of antibody staining. This consisted of extraction of unstained portions of the sarcomere with 0.6 M potassium iodide, 10-4 M adenosine triphosphate solution. Stained regions of the sarcomere remained intact because of insolubility of the combined antigen and antibody.

scholarly journals An Effective Method of Preparing Sections of Bacillus polymyxa Sporangia and Spores for Electron Microscopy

1960 ◽  
Vol 7 (2) ◽  
pp. 373-376 ◽  
Author(s):  
Pauline E. Holbert
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Electron Micrographs ◽  
Epoxy Resins ◽  
Bacillus Polymyxa ◽  
Thin Sections ◽  
Similar Images ◽  
Embedding Medium ◽  
First Time ◽  
Diamond Knife

Bacillus polymyxa sporangia and spores were prepared for examination in the electron microscope by methods whose critical features were apparently: judicious use of vacuum, to encourage complete penetration of the embedding medium; the use of epoxy resins as embedding media; and cutting of the thin sections with a diamond knife. Electron micrographs of material prepared in this manner exhibit undeformed sporangial sections. Some of the structures revealed have been shown before, though perhaps less distinctly; other structures are revealed here for the first time. While this single study does not pretend to elucidate all the complexities of sporulation in bacteria, these and similar images should make this possible, and some mention of the preparatory techniques that lead to them seems advisable at this time.

The Biological Electron Microscope Facility, University of Hawai‘I at Mānoa: a Shared Microscopy Resource in the Pacific

1997 ◽  
Vol 3 (S2) ◽  
pp. 285-286
Author(s):  
T.M. Carvalho ◽  
M.F. Dunlap ◽  
R.D. Allen
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Research University ◽  
State Of The Art ◽  
Research Unit ◽  
The Past ◽  
The Pacific ◽  
University Of Hawai'i ◽  
The University ◽  
And Training

The Biological Electron Microscope Facility (BEMF) at the University of Hawai‘i at Mānoa (UHM) is located 2400 miles over water from the next nearest research university. BEMF is a multi-user core facility, administered by the Pacific Biomedical Research Center (PBRC), an organized research unit at the UHM. The mission of the BEMF is to provide state-of-the-art instrumentation, services and training for electron microscopy to the biomedical and biological researchers in Hawai‘i and the Pacific region. The BEMF was established in 1984 under the direction of Dr. Richard D. Allen, and has since grown steadily in its instrumentation, expertise, and use. In the past 5 years it has served researchers from over 50 laboratories in PBRC and the colleges of Natural Sciences, Tropical Agriculture and Human Resources, Engineering, Medicine, and Ocean and Earth Sciences and Technology, as well as visiting investigators from other Hawai‘i, mainland and foreign institutions.The BEMF has a full line of instrumentation for conventional transmission and field emission scanning electron microscopy as well as a complete line of instruments for cryoelectron microscopy.

scholarly journals ELECTRON MICROSCOPY OF SPORES OF BACILLUS MEGATERIUM WITH SPECIAL REFERENCE TO THE EFFECTS OF FIXATION AND THIN SECTIONING

1962 ◽  
Vol 13 (3) ◽  
pp. 423-435 ◽  
Author(s):  
L. J. Rode ◽  
C. Willard Lewis ◽  
J. W. Foster
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Special Reference ◽  
Bacillus Megaterium ◽  
Normal State ◽  
Dipicolinic Acid ◽  
Indirect Evidence ◽  
Outer Edge ◽  
Thin Sectioning ◽  
Resting Spores

Resting spores of Bacillus megaterium appear uniformly opaque and undifferentiated under the electron microscope. Germinated spores and spores which have lost their dipicolinic acid underwent characteristic changes in structure. Spores fixed with KMnO4 lose their dipicolinic acid. Spores fixed with OsO4 under certain conditions retain their dipicolinic acid. When conventional sectioning procedures are used with either method of fixation, abnormal spore structure is produced as a result of the solution of cellular constitutents. Dry sections of unfixed spores embedded in methacrylate reveal the spore structure in a more normal state. Indirect evidence has been obtained for the existence of a penetration barrier at or near the outer edge of the cortex.

Thin sectioning for cryo transmission electron microscopy (cryo TEM)

1986 ◽  
Vol 44 ◽  
pp. 102-103
Author(s):  
A.W. McDowall ◽  
J.M. Smith ◽  
J. Dubochet
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Electron Microscope ◽  
Structural Integrity ◽  
Specimen Preparation ◽  
Structural Alterations ◽  
Whole Cells ◽  
Transmission Electron ◽  
Thin Sectioning

Processing whole cells and tissues for conventional TEM is known to cause structural alterations. Much effort has been devoted, therefore, to developing techniques which avoid specimen preparation artefacts. Recently, research using a cryo-electron microscope has shown that biological suspensions embedded in vitreous ice retain their structural integrity, and when compared with conventionally prepared TEM specimens, are free from many of the classical artefacts. In order to extend the advantage of cryo TEM to whole cells and tissues, we have developed a method of thin sectioning vitrified material.

scholarly journals Araldite as an Embedding Medium for Electron Microscopy

1958 ◽  
Vol 4 (2) ◽  
pp. 191-194 ◽  
Author(s):  
Audrey M. Glauert ◽  
R. H. Glauert
Keyword(s):  
Electron Microscopy ◽  
Fine Structure ◽  
Epoxy Resin ◽  
Epoxy Resins ◽  
Thin Sectioning ◽  
Embedding Medium ◽  
Final Block

Epoxy resins are suitable media for embedding for electron microscopy, as they set uniformly with virtually no shrinkage. A mixture of araldite epoxy resins has been developed which is soluble in ethanol, and which yields a block of the required hardness for thin sectioning. The critical modifications to the conventional mixtures are the choice of a plasticized resin in conjunction with an aliphatic anhydride as the hardener. The hardness of the final block can be varied by incorporating additional plasticizer, and the rate of setting can be controlled by the use of an amine accelerator. The properties of the araldite mixture can be varied quite widely by adjusting the proportions of the various constituents. The procedure for embedding biological specimens is similar to that employed with methacrylates, although longer soaking times are recommended to ensure the complete penetration of the more viscous epoxy resin. An improvement in the preservation of the fine structure of a variety of specimens has already been reported, and a typical electron microgram illustrates the present paper.

Electron microscopy in the community hospital

1987 ◽  
Vol 45 ◽  
pp. 610-611
Author(s):  
Gary S. Pearl
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Community Hospital ◽  
Easy Access ◽  
Teaching Program ◽  
The Past ◽  
Common Misconception ◽  
Ultrastructural Findings

A common misconception exists that a diagnostic electron microscopy laboratory can not succeed fiscally in a community hospital. This has not been the case in our institution. An electron microscopy laboratory was established in 1982 and the initial investment by the hospital was recovered within four years. Billings have remained relatively constant during the past several years. In addition to financial considerations, the laboratory has been beneficial diagnostically, allowing for rapid, easy access to this valuable tool. In fact, the presence of an electron microscope in the hospital has probably encouraged further utilization of this technology. The electron microscopy laboratory has also been an asset in our teaching program, not only for the demonstration of the ultrastructural findings themselves, but also for the consultative material sent from other institutions. Finally, the laboratory has opened up research possibilities not otherwise available.

Immune electron microscopy of haemocyanins from two southern african whelks

1978 ◽  
Vol 36 (2) ◽  
pp. 158-159
Author(s):  
Linda M. Stannard ◽  
Margaret Lennon
Keyword(s):  
Electron Microscopy ◽  
Electron Microscope ◽  
Outer Ring ◽  
Immune Electron Microscopy ◽  
Intertidal Zones ◽  
Central Belt ◽  
Cape Peninsula ◽  
Circular Contour

Burnupena cincta and Fusus verruculatus are two whelks which inhabit the intertidal zones of the Cape Peninsula shore. Their respiratory pigments, or haemocyanins, are morphologically similar in structure (Figs. 1 and 2) and appear in the electron microscope as short cylindrical rods about 34 nm in diameter and 36 nm high. Viewed side-on the molecules show regular banding suggesting a structure composed of six equidistant rings of sub-units. Occasionally the particles have the appearance of possessing a central “belt” in the position of the 3rd and 4th rows of sub-units. End-on views of the haemocyanin molecules show a circular contour with a dense outer ring and a less dense inner ring in which 10 definite sub-units may frequently be distinguished. A number of molecules display an extra central inner component which appears either as a diffuse plug or as a discrete ring-shaped core ± 8 nm in diameter.

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