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Related Concept Videos

Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...

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Related Experiment Video

Updated: Jun 25, 2026

Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data
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Relative Quantification of Cellular Sections with Molecular Depth Profiling ToF-SIMS Imaging.

M E Kurczy1, Joseph Kozole, S A Parry

  • 1Department of Chemistry, Penn State University, 104 Chemistry Building, University Park, PA 16802.

Applied Surface Science
|February 28, 2009
PubMed
Summary
This summary is machine-generated.

Secondary ion mass spectrometry (SIMS) imaging quantifies lipid differences between cell membranes and cytoplasm. This method enables comparative lipid analysis in treated versus control cells using lipophilic dyes for depth profiling.

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Area of Science:

  • Cell biology
  • Biophysics
  • Analytical chemistry

Background:

  • Lipid distribution within single cells is crucial for cellular function.
  • Quantifying lipid differences between cellular compartments (plasma membrane and cytoplasm) presents analytical challenges.
  • Existing methods may lack the spatial resolution or sensitivity for detailed lipid profiling.

Purpose of the Study:

  • To develop and demonstrate a SIMS imaging approach for quantifying relative lipid differences between the plasma membrane and cytoplasm of single cells.
  • To utilize lipophilic dyes as markers for cell identification and depth profiling within SIMS analysis.
  • To establish a method for comparing lipid headgroup amounts in treated versus control cells.

Main Methods:

  • Time-of-Flight Secondary Ion Mass Spectrometry (ToF-SIMS) imaging with a C(60)(+) ion source.
  • Labeling of cells with lipophilic dyes for membrane identification and depth analysis.
  • Cryo-fracturing of cells to expose internal structures (plasma membrane and cytoplasm).
  • Analysis of multiple cellular sections following a single fracture event.

Main Results:

  • Successful quantification of relative lipid amounts between the plasma membrane and cytoplasm in single cells.
  • Demonstration of lipophilic dyes as effective markers for cell population identification and membrane depth profiling.
  • Validation of the ability to compare specific lipid headgroup quantities in treated cells versus control cells.
  • Showcased the capacity to analyze multiple cellular sections from a single fracture.

Conclusions:

  • SIMS imaging provides a powerful tool for quantitative lipid analysis within distinct cellular compartments.
  • The developed method allows for comparative lipid profiling between different cell populations or treatment conditions.
  • This approach offers a foundation for detailed investigations into cellular lipidomics and membrane biophysics.