Identification of cellular sections with imaging mass spectrometry following freeze fracture

Thomas P. Roddy, Donald M. Cannon, Sara G. Ostrowski, Nicholas Winograd, Andrew G. Ewing

Research output: Contribution to journalArticle

73 Citations (Scopus)

Abstract

Freeze-fracture techniques have been used to maintain chemical heterogeneity of frozen-hydrated mammalian cells for static TOF-SIMS imaging. The effects the fracture plane has on scanning electron microscopy and dynamic SIMS images of cells have been studied, but the implications this preparation method has on static SIMS have not been addressed to date. Interestingly, the chemical specificity and surface sensitivity of TOF-SIMS have allowed the identification of unique sections of rat pheochromocytoma cells exposed to the sample surface during freeze fracture. Using the extensive chemical information of the fractured surface, cellular sections have been determined using TOF-SIMS images of water, sodium, potassium, hydrocarbons, phosphocholine, and DiI, a fluorescent dye that remains in the outer leaflet of the cell membrane. Higher amounts of potassium have been imaged inside a cell versus the surrounding matrix in a cross-fractured cell. In other fractures exposing the cell membrane, phosphocholine and DiI have been imaged on the outer leaflet of the cell membrane, while phosphocholine alone has been imaged on the inner leaflet. In this paper, we discuss how imaging mass spectrometry is used to uniquely distinguish three possible sections of cells obtained during freeze fracture. The identification of these sections is important in choosing cells with a region of interest, like the cell membrane, exposed to the surface for a more thorough investigation with imaging static TOF-SIMS.

Original languageEnglish (US)
Pages (from-to)4020-4026
Number of pages7
JournalAnalytical Chemistry
Volume74
Issue number16
DOIs
StatePublished - Aug 15 2002

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Secondary ion mass spectrometry
Mass spectrometry
Cell membranes
Imaging techniques
Phosphorylcholine
Potassium
Hydrocarbons
Fluorescent Dyes
Rats
Sodium
Cells
Scanning electron microscopy
Water

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Roddy, Thomas P. ; Cannon, Donald M. ; Ostrowski, Sara G. ; Winograd, Nicholas ; Ewing, Andrew G. / Identification of cellular sections with imaging mass spectrometry following freeze fracture. In: Analytical Chemistry. 2002 ; Vol. 74, No. 16. pp. 4020-4026.
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Identification of cellular sections with imaging mass spectrometry following freeze fracture. / Roddy, Thomas P.; Cannon, Donald M.; Ostrowski, Sara G.; Winograd, Nicholas; Ewing, Andrew G.

In: Analytical Chemistry, Vol. 74, No. 16, 15.08.2002, p. 4020-4026.

Research output: Contribution to journalArticle

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