Immunogold labeling of bacterial cells for transmission electron microscopy (TEM) v1 (protocols.io.mv4c68w)

protocols.io ◽  
2018 ◽  
Author(s):  
Jessica Sacher
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Immunogold Labeling ◽  
Bacterial Cells ◽  
Transmission Electron

Molecular and atomic analysis of uranium complexes formed by three eco-types of Acidithiobacillus ferrooxidans

2002 ◽  
Vol 30 (4) ◽  
pp. 669-672 ◽  
Author(s):  
M. Merroun ◽  
C. Hennig ◽  
A. Rossberg ◽  
G. Geipel ◽  
T. Reich ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Acidithiobacillus Ferrooxidans ◽  
Energy Dispersive ◽  
Bacterial Cells ◽  
X Ray ◽  
Uranium Complexes ◽  
Transmission Electron ◽  
Polyphosphate Bodies ◽  
Atomic Analysis

A combination of EXAFS, transmission electron microscopy and energy-dispersive X-ray was used to conduct a molecular and atomic analysis of the uranium complexes formed by Acidithiobacillus ferrooxidans. The results demonstrate that this bacterium accumulates uranium as phosphate compounds. We suggest that at toxic levels when the uranium enters the bacterial cells, A. ferrooxidans can detoxify and efflux this metal by a process in which its polyphosphate bodies are involved.

Determination of Bacterial Cell Dry Mass by Transmission Electron Microscopy and Densitometric Image Analysis

1998 ◽  
Vol 64 (2) ◽  
pp. 688-694 ◽  
Author(s):  
M. Loferer-Krößbacher ◽  
J. Klima ◽  
R. Psenner
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Image Analysis ◽  
Bacterial Biomass ◽  
Dry Weight ◽  
Dry Mass ◽  
Bacterial Cells ◽  
E Coli ◽  
Transmission Electron ◽  
Bacterial Assemblages

ABSTRACT We applied transmission electron microscopy and densitometric image analysis to measure the cell volume (V) and dry weight (DW) of single bacterial cells. The system was applied to measure the DW ofEscherichia coli DSM 613 at different growth phases and of natural bacterial assemblages of two lakes, Piburger See and Gossenköllesee. We found a functional allometric relationship between DW (in femtograms) and V (in cubic micrometers) of bacteria (DW = 435 · V 0.86); i.e., smaller bacteria had a higher ratio of DW to V than larger cells. The measured DW of E. coli cells ranged from 83 to 1,172 fg, and V ranged from 0.1 to 3.5 μm3(n = 678). Bacterial cells from Piburger See and Gossenköllesee (n = 465) had DWs from 3 fg (V = 0.003 μm3) to 1,177 fg (V = 3.5 μm3). Between 40 and 50% of the cells had a DW of less than 20 fg. By assuming that carbon comprises 50% of the DW, the ratio of carbon content to Vof individual cells varied from 466 fg of C μm−3 forVs of 0.001 to 0.01 μm3 to 397 fg of C μm−3 (0.01 to 0.1 μm3) and 288 fg of C μm−3 (0.1 to 1 μm3). Exponentially growing and stationary cells of E. coli DSM 613 showed conversion factors of 254 fg of C μm−3 (0.1 to 1 μm3) and 211 fg of C μm−3 (1 to 4 μm3), respectively. Our data suggest that bacterial biomass in aquatic environments is higher and more variable than previously assumed from volume-based measurements.

Hexavalent chromium uptake by an Arthrobacter species: Electron microscope studies

1984 ◽  
Vol 42 ◽  
pp. 680-681
Author(s):  
R. Bhatnagar ◽  
R. N. Coleman
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Present Report ◽  
Bacterial Cells ◽  
Chromium Vi ◽  
Transmission Electron ◽  
Bacterial Removal ◽  
Scanning Transmission ◽  
Arthrobacter Species ◽  
Micro Organisms

Many micro-organisms are capable of taking up and accumulating specific metal ions from their environment. As a part of a study to evaluate bacterial removal of chromium from waste liquid streams, bacterial isolate (#96) identified as an Arthrobacter sp. was selected from 362 isolates and was reported to exhibit high tolerance to and uptake of chromium VI as determined by chemical methods.2 The present report describes fine structural studies of #96 with Transmission Electron Microscopy (TEM) and Scanning Transmission Electron Microscopy (STEM) to further understand the location of chromium accumulation and uptake. Attempts have been made to localize chromium on/in bacterial cells using Energy-Dispersive X-ray Microanalysis (EDX) and elemental mapping.

Immunogold labeling of HTLV-III/LAV in H9 cells studied by transmission electron microscopy

1986 ◽  
Vol 13 (3) ◽  
pp. 265-269 ◽  
Author(s):  
D. Pekovic ◽  
S. Garzon ◽  
H. Strykowski ◽  
D. Ajdukovic ◽  
M. Gornitsky ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Immunogold Labeling ◽  
Transmission Electron

scholarly journals Pitfalls of immunogold labeling: analysis by light microscopy, transmission electron microscopy, and photoelectron microscopy.

1987 ◽  
Vol 35 (8) ◽  
pp. 843-853 ◽  
Author(s):  
G B Birrell ◽  
K K Hedberg ◽  
O H Griffith
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Light Microscopy ◽  
Tannic Acid ◽  
Cultured Cells ◽  
Immunogold Labeling ◽  
Fish Gelatin ◽  
Nonspecific Binding ◽  
Gold Particles ◽  
Transmission Electron

The immunogold method is widely used to localize, identify, and distinguish cellular antigens. There are, however, some pitfalls that can lead to nonspecific binding, particularly in cytoskeletal studies with gold probes prepared from small gold particles. We present a list of suggestions for minimizing nonspecific binding, with particular attention to two problems identified in this study. First, we find that the method used to prepare the colloidal gold particles affects the degree of nonspecific binding. Second, the standard BSA-stabilized small gold probes evidently possess exposed regions that bind to the proteins of cytoskeletal preparations. This was investigated in whole-mount cytoskeletal preparations of cultured cells by use of light microscopy, transmission electron microscopy, and photoelectron microscopy of silver-enhanced specimens. Gold probes were made from approximately 5-nm particles generated by reduction of HAuCl4 with three different reducing agents: white phosphorus, sodium borohydride, and citrate-tannic acid. All three preparations stabilized in the conventional way showed significant levels of nonspecific binding, which was highest with citrate-tannic acid. This problem was largely solved with all three types of probes by including fish gelatin in the probe buffer, by substituting fish gelatin for the BSA stabilizer used to prepare the probes, or by pre-adsorption methods. Application of these techniques resulted in clear immunogold labeling patterns with minimal nonspecific background.

Ultrastructure and immunogold labeling of cytoplasmic fibrous bundles in tobacco leaf cells infected with tobacco vein mottling virus

1992 ◽  
Vol 50 (1) ◽  
pp. 676-677
Author(s):  
E. D. Ammar ◽  
D. W. Thombury ◽  
T. P. Pirone
Keyword(s):  
Electron Microscopy ◽  
Transmission Electron Microscopy ◽  
Immunogold Labeling ◽  
Infected Leaf ◽  
Transmission Electron ◽  
Aphid Vectors ◽  
Cylindrical Inclusions ◽  
Tobacco Vein Mottling Virus ◽  
Unknown Composition ◽  
Infected Tobacco

Tobacco vein mottling virus (TVMV) is a potyvirus, with flexuous filamentous particles ca. 765 nm long and 12-13 nm in diameter. TVMV is transmitted by sap inoculation and by aphid vectors. Cylindrical inclusions characteristic of infections with potyviruses have been found in the cytoplasm of TVMV-infected tobacco leaves and protoplasts, and the presence of fibrous inclusions of unknown composition in the cytoplasm and nuclei of TVMV-infected tobacco has been reported. Here, we report the results of transmission electron microscopy (TEM) and immunogold labeling of cytoplasmic fibrous bundles found in TVMV-infected leaf cells, providing evidence that these bundles are composed of TVMV virions and/or coat protein.

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