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Title: Mounting of Escherichia coli spheroplasts for AFM imaging.

Abstract

The cytoplasmic membrane of Escherichia coli (E. coli) is the location of numerous, chemically specific transporters and recognition elements. Investigation of this membrane in vivo by atomic force microscopy (AFM) requires removal of the cell wall and stable immobilization of the spheroplast. AFM images demonstrate that spheroplasts can be secured with warm gelatin applied to the mica substrate just before the addition of a spheroplast suspension. The resulting preparation can be repeatedly imaged by AFM over the course of several hours. Confocal fluorescence imaging confirms the association of the spheroplasts with the gelatin layer. Gelatin molecules are known to reorder into a network after heating. Entrapment within this gelatin network is believed to be responsible for the immobilization of spheroplasts on mica.

Authors:
 [1];  [1];  [1];  [1]
  1. ORNL
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
989679
DOE Contract Number:
DE-AC05-00OR22725
Resource Type:
Journal Article
Resource Relation:
Journal Name: Ultramicroscopy; Journal Volume: 105; Journal Issue: 1-4
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ATOMIC FORCE MICROSCOPY; ESCHERICHIA COLI; SAMPLE PREPARATION; FLUORESCENCE; IN VIVO; MEMBRANES; MICA; SUBSTRATES

Citation Formats

Sullivan, Claretta J, Morrell-Falvey, Jennifer L, Allison, David P, and Doktycz, Mitchel John. Mounting of Escherichia coli spheroplasts for AFM imaging.. United States: N. p., 2005. Web. doi:10.1016/j.ultramic.2005.06.023.
Sullivan, Claretta J, Morrell-Falvey, Jennifer L, Allison, David P, & Doktycz, Mitchel John. Mounting of Escherichia coli spheroplasts for AFM imaging.. United States. doi:10.1016/j.ultramic.2005.06.023.
Sullivan, Claretta J, Morrell-Falvey, Jennifer L, Allison, David P, and Doktycz, Mitchel John. Tue . "Mounting of Escherichia coli spheroplasts for AFM imaging.". United States. doi:10.1016/j.ultramic.2005.06.023.
@article{osti_989679,
title = {Mounting of Escherichia coli spheroplasts for AFM imaging.},
author = {Sullivan, Claretta J and Morrell-Falvey, Jennifer L and Allison, David P and Doktycz, Mitchel John},
abstractNote = {The cytoplasmic membrane of Escherichia coli (E. coli) is the location of numerous, chemically specific transporters and recognition elements. Investigation of this membrane in vivo by atomic force microscopy (AFM) requires removal of the cell wall and stable immobilization of the spheroplast. AFM images demonstrate that spheroplasts can be secured with warm gelatin applied to the mica substrate just before the addition of a spheroplast suspension. The resulting preparation can be repeatedly imaged by AFM over the course of several hours. Confocal fluorescence imaging confirms the association of the spheroplasts with the gelatin layer. Gelatin molecules are known to reorder into a network after heating. Entrapment within this gelatin network is believed to be responsible for the immobilization of spheroplasts on mica.},
doi = {10.1016/j.ultramic.2005.06.023},
journal = {Ultramicroscopy},
number = 1-4,
volume = 105,
place = {United States},
year = {Tue Nov 01 00:00:00 EST 2005},
month = {Tue Nov 01 00:00:00 EST 2005}
}
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