Time-of-flight secondary ion mass spectrometry imaging of subcellular lipid heterogeneity: Poisson counting and spatial resolution

Paul D. Piehowski, Angel M. Davey, Michael E. Kurczy, Erin D. Sheets, Nicholas Winograd, Andrew G. Ewing, Michael L. Heien

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Mass spectrometric imaging is a powerful tool to interrogate biological complexity. One such technique, time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging, has been successfully utilized for subcellular imaging of cell membrane components. In order for this technique to provide insight into biological processes, it is critical to characterize the figures of merit. Because a SIMS instrument counts individual events, the precision of the measurement is controlled by counting statistics. As the analysis area decreases, the number of molecules available for analysis diminishes. This becomes critical when imaging subcellular features; it limits the information obtainable, resulting in images with only a few counts of interest per pixel. Many features observed in low intensity images are artifacts of counting statistics, making validation of these features crucial to arriving at accurate conclusions. With TOF-SIMS imaging, the experimentally attainable spatial resolution is a function of the molecule of interest, sample matrix, concentration, primary ion, instrument transmission, and spot size of the primary ion beam. A model, based on Poisson statistics, has been developed to validate SIMS imaging data when signal is limited. This model can be used to estimate the effective spatial resolution and limits of detection prior to analysis, making it a powerful tool for tailoring future investigations. In addition, the model allows comparison of pixel-to-pixel intensity and can be used to validate the significance of observed image features. The implications and capabilities of the model are demonstrated by imaging the cell membrane of resting RBL-2H3 mast cells.

Original languageEnglish (US)
Pages (from-to)5593-5602
Number of pages10
JournalAnalytical chemistry
Volume81
Issue number14
DOIs
StatePublished - Jul 15 2009

Fingerprint

Secondary ion mass spectrometry
Lipids
Imaging techniques
Pixels
Statistics
Cell membranes
Molecules
Ion beams
Ions

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Piehowski, Paul D. ; Davey, Angel M. ; Kurczy, Michael E. ; Sheets, Erin D. ; Winograd, Nicholas ; Ewing, Andrew G. ; Heien, Michael L. / Time-of-flight secondary ion mass spectrometry imaging of subcellular lipid heterogeneity : Poisson counting and spatial resolution. In: Analytical chemistry. 2009 ; Vol. 81, No. 14. pp. 5593-5602.
@article{05e3a9d413b74d4ebdc3796d296d2ed4,
title = "Time-of-flight secondary ion mass spectrometry imaging of subcellular lipid heterogeneity: Poisson counting and spatial resolution",
abstract = "Mass spectrometric imaging is a powerful tool to interrogate biological complexity. One such technique, time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging, has been successfully utilized for subcellular imaging of cell membrane components. In order for this technique to provide insight into biological processes, it is critical to characterize the figures of merit. Because a SIMS instrument counts individual events, the precision of the measurement is controlled by counting statistics. As the analysis area decreases, the number of molecules available for analysis diminishes. This becomes critical when imaging subcellular features; it limits the information obtainable, resulting in images with only a few counts of interest per pixel. Many features observed in low intensity images are artifacts of counting statistics, making validation of these features crucial to arriving at accurate conclusions. With TOF-SIMS imaging, the experimentally attainable spatial resolution is a function of the molecule of interest, sample matrix, concentration, primary ion, instrument transmission, and spot size of the primary ion beam. A model, based on Poisson statistics, has been developed to validate SIMS imaging data when signal is limited. This model can be used to estimate the effective spatial resolution and limits of detection prior to analysis, making it a powerful tool for tailoring future investigations. In addition, the model allows comparison of pixel-to-pixel intensity and can be used to validate the significance of observed image features. The implications and capabilities of the model are demonstrated by imaging the cell membrane of resting RBL-2H3 mast cells.",
author = "Piehowski, {Paul D.} and Davey, {Angel M.} and Kurczy, {Michael E.} and Sheets, {Erin D.} and Nicholas Winograd and Ewing, {Andrew G.} and Heien, {Michael L.}",
year = "2009",
month = "7",
day = "15",
doi = "10.1021/ac901065s",
language = "English (US)",
volume = "81",
pages = "5593--5602",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "14",

}

Time-of-flight secondary ion mass spectrometry imaging of subcellular lipid heterogeneity : Poisson counting and spatial resolution. / Piehowski, Paul D.; Davey, Angel M.; Kurczy, Michael E.; Sheets, Erin D.; Winograd, Nicholas; Ewing, Andrew G.; Heien, Michael L.

In: Analytical chemistry, Vol. 81, No. 14, 15.07.2009, p. 5593-5602.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Time-of-flight secondary ion mass spectrometry imaging of subcellular lipid heterogeneity

T2 - Poisson counting and spatial resolution

AU - Piehowski, Paul D.

AU - Davey, Angel M.

AU - Kurczy, Michael E.

AU - Sheets, Erin D.

AU - Winograd, Nicholas

AU - Ewing, Andrew G.

AU - Heien, Michael L.

PY - 2009/7/15

Y1 - 2009/7/15

N2 - Mass spectrometric imaging is a powerful tool to interrogate biological complexity. One such technique, time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging, has been successfully utilized for subcellular imaging of cell membrane components. In order for this technique to provide insight into biological processes, it is critical to characterize the figures of merit. Because a SIMS instrument counts individual events, the precision of the measurement is controlled by counting statistics. As the analysis area decreases, the number of molecules available for analysis diminishes. This becomes critical when imaging subcellular features; it limits the information obtainable, resulting in images with only a few counts of interest per pixel. Many features observed in low intensity images are artifacts of counting statistics, making validation of these features crucial to arriving at accurate conclusions. With TOF-SIMS imaging, the experimentally attainable spatial resolution is a function of the molecule of interest, sample matrix, concentration, primary ion, instrument transmission, and spot size of the primary ion beam. A model, based on Poisson statistics, has been developed to validate SIMS imaging data when signal is limited. This model can be used to estimate the effective spatial resolution and limits of detection prior to analysis, making it a powerful tool for tailoring future investigations. In addition, the model allows comparison of pixel-to-pixel intensity and can be used to validate the significance of observed image features. The implications and capabilities of the model are demonstrated by imaging the cell membrane of resting RBL-2H3 mast cells.

AB - Mass spectrometric imaging is a powerful tool to interrogate biological complexity. One such technique, time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging, has been successfully utilized for subcellular imaging of cell membrane components. In order for this technique to provide insight into biological processes, it is critical to characterize the figures of merit. Because a SIMS instrument counts individual events, the precision of the measurement is controlled by counting statistics. As the analysis area decreases, the number of molecules available for analysis diminishes. This becomes critical when imaging subcellular features; it limits the information obtainable, resulting in images with only a few counts of interest per pixel. Many features observed in low intensity images are artifacts of counting statistics, making validation of these features crucial to arriving at accurate conclusions. With TOF-SIMS imaging, the experimentally attainable spatial resolution is a function of the molecule of interest, sample matrix, concentration, primary ion, instrument transmission, and spot size of the primary ion beam. A model, based on Poisson statistics, has been developed to validate SIMS imaging data when signal is limited. This model can be used to estimate the effective spatial resolution and limits of detection prior to analysis, making it a powerful tool for tailoring future investigations. In addition, the model allows comparison of pixel-to-pixel intensity and can be used to validate the significance of observed image features. The implications and capabilities of the model are demonstrated by imaging the cell membrane of resting RBL-2H3 mast cells.

UR - http://www.scopus.com/inward/record.url?scp=67650725816&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=67650725816&partnerID=8YFLogxK

U2 - 10.1021/ac901065s

DO - 10.1021/ac901065s

M3 - Article

C2 - 19530687

AN - SCOPUS:67650725816

VL - 81

SP - 5593

EP - 5602

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 14

ER -