Improved transmission electron microscopy (TEM) of cultured cells through a "floating sheet" method

James R. Arnold, Paul J. Boor

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Cultured cells provide isolated systems for both biochemical and morphological studies. Previous methods of processing cell culture specimens for electron microscopy (EM) have been limited to sectioning either a monolayer or centrifuged cell suspensions which are not morphologically intact. In our improved method, N-butylglycidyl ether is added to cell cultures (2-5 min with agitation) following in situ fixation (3.0% glutaraldehyde in 0.1 M Pipes, pH 7.2, for 20 min, osmium tetraoxide 4% for 20 min). A thin pliable "sheet" of cells floats free from the plastic culture device and can be manipulated (centrifuged or folded) to obtain a vast number of morphologically intact cells for examination. We have examined several cell types (vascular smooth muscle, lung, liver, and endothelial cells) grown in two types of plastic culture flasks (Nunc and Falcon). This new method provides excellent EM morphology, maximizes the number of cells examined, and allows determination of cell orientation since a remnant of the dissolved flask remains loosely bound to the bottom of the cells.

Original languageEnglish (US)
Pages (from-to)30-36
Number of pages7
JournalJournal of Ultrastructure Research and Molecular Structure Research
Volume94
Issue number1
DOIs
StatePublished - 1986

Fingerprint

Transmission Electron Microscopy
Cultured Cells
Plastics
Electron Microscopy
Cell Culture Techniques
Osmium
Glutaral
Vascular Smooth Muscle
Ether
Suspensions
Endothelial Cells
Cell Count
Equipment and Supplies
Lung
Liver

ASJC Scopus subject areas

  • Molecular Biology
  • Anatomy

Cite this

Improved transmission electron microscopy (TEM) of cultured cells through a "floating sheet" method. / Arnold, James R.; Boor, Paul J.

In: Journal of Ultrastructure Research and Molecular Structure Research, Vol. 94, No. 1, 1986, p. 30-36.

Research output: Contribution to journalArticle

@article{b4dbfa2ac1e84ddaa49e0818396e9c65,
title = "Improved transmission electron microscopy (TEM) of cultured cells through a {"}floating sheet{"} method",
abstract = "Cultured cells provide isolated systems for both biochemical and morphological studies. Previous methods of processing cell culture specimens for electron microscopy (EM) have been limited to sectioning either a monolayer or centrifuged cell suspensions which are not morphologically intact. In our improved method, N-butylglycidyl ether is added to cell cultures (2-5 min with agitation) following in situ fixation (3.0{\%} glutaraldehyde in 0.1 M Pipes, pH 7.2, for 20 min, osmium tetraoxide 4{\%} for 20 min). A thin pliable {"}sheet{"} of cells floats free from the plastic culture device and can be manipulated (centrifuged or folded) to obtain a vast number of morphologically intact cells for examination. We have examined several cell types (vascular smooth muscle, lung, liver, and endothelial cells) grown in two types of plastic culture flasks (Nunc and Falcon). This new method provides excellent EM morphology, maximizes the number of cells examined, and allows determination of cell orientation since a remnant of the dissolved flask remains loosely bound to the bottom of the cells.",
author = "Arnold, {James R.} and Boor, {Paul J.}",
year = "1986",
doi = "10.1016/0889-1605(86)90049-2",
language = "English (US)",
volume = "94",
pages = "30--36",
journal = "Journal of Structural Biology",
issn = "1047-8477",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Improved transmission electron microscopy (TEM) of cultured cells through a "floating sheet" method

AU - Arnold, James R.

AU - Boor, Paul J.

PY - 1986

Y1 - 1986

N2 - Cultured cells provide isolated systems for both biochemical and morphological studies. Previous methods of processing cell culture specimens for electron microscopy (EM) have been limited to sectioning either a monolayer or centrifuged cell suspensions which are not morphologically intact. In our improved method, N-butylglycidyl ether is added to cell cultures (2-5 min with agitation) following in situ fixation (3.0% glutaraldehyde in 0.1 M Pipes, pH 7.2, for 20 min, osmium tetraoxide 4% for 20 min). A thin pliable "sheet" of cells floats free from the plastic culture device and can be manipulated (centrifuged or folded) to obtain a vast number of morphologically intact cells for examination. We have examined several cell types (vascular smooth muscle, lung, liver, and endothelial cells) grown in two types of plastic culture flasks (Nunc and Falcon). This new method provides excellent EM morphology, maximizes the number of cells examined, and allows determination of cell orientation since a remnant of the dissolved flask remains loosely bound to the bottom of the cells.

AB - Cultured cells provide isolated systems for both biochemical and morphological studies. Previous methods of processing cell culture specimens for electron microscopy (EM) have been limited to sectioning either a monolayer or centrifuged cell suspensions which are not morphologically intact. In our improved method, N-butylglycidyl ether is added to cell cultures (2-5 min with agitation) following in situ fixation (3.0% glutaraldehyde in 0.1 M Pipes, pH 7.2, for 20 min, osmium tetraoxide 4% for 20 min). A thin pliable "sheet" of cells floats free from the plastic culture device and can be manipulated (centrifuged or folded) to obtain a vast number of morphologically intact cells for examination. We have examined several cell types (vascular smooth muscle, lung, liver, and endothelial cells) grown in two types of plastic culture flasks (Nunc and Falcon). This new method provides excellent EM morphology, maximizes the number of cells examined, and allows determination of cell orientation since a remnant of the dissolved flask remains loosely bound to the bottom of the cells.

UR - http://www.scopus.com/inward/record.url?scp=0023017240&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023017240&partnerID=8YFLogxK

U2 - 10.1016/0889-1605(86)90049-2

DO - 10.1016/0889-1605(86)90049-2

M3 - Article

VL - 94

SP - 30

EP - 36

JO - Journal of Structural Biology

JF - Journal of Structural Biology

SN - 1047-8477

IS - 1

ER -