Mounting of Escherichia coli spheroplasts for AFM imaging

C. J. Sullivan; J. L. Morrell; D. P. Allison; M. J. Doktycz

doi:10.1016/j.ultramic.2005.06.023

Mounting of Escherichia coli spheroplasts for AFM imaging

C. J. Sullivan, J. L. Morrell, D. P. Allison, M. J. Doktycz

Research output: Contribution to journal › Article › peer-review

27 Scopus citations

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.

Original language	English
Pages (from-to)	96-102
Number of pages	7
Journal	Ultramicroscopy
Volume	105
Issue number	1-4
DOIs	https://doi.org/10.1016/j.ultramic.2005.06.023
State	Published - Nov 2005

Funding

The authors wish to thank Dr. Dale Pelletier for generously donating the GFP expressing E. coli. This research was sponsored by the Office of Biological and Environmental Research, US Department of Energy. Oak Ridge National Laboratory is managed by UT-Battelle, LLC, for the US Department of Energy under Contract no. DE-AC05-00OR22725.

Keywords

Atomic force microscopy
Bacteria
Cytoplasmic membrane
Escherichia coli
Gelatin
Immobilization
Live cell imaging
Macmode
Spheroplasts

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10.1016/j.ultramic.2005.06.023

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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.",

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AU - Morrell, J. L.

AU - Allison, D. P.

AU - Doktycz, M. J.

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AB - 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.

KW - Atomic force microscopy

KW - Bacteria

KW - Cytoplasmic membrane

KW - Escherichia coli

KW - Gelatin

KW - Immobilization

KW - Live cell imaging

KW - Macmode

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Mounting of Escherichia coli spheroplasts for AFM imaging

Abstract

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