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Updated: Jan 21, 2026

Multi-color Localization Microscopy of Single Membrane Proteins in Organelles of Live Mammalian Cells
Published on: June 30, 2018
Toward Single-Organelle Lipidomics in Live Cells
Adrian Lita1, Andrey N Kuzmin2,3, Artem Pliss2,3
1Neuro-Oncology Branch, Center for Cancer Research , National Cancer Institute, National Institutes of Health , Bethesda , Maryland 20892 , United States.
This study introduces a novel micro-Raman assay for single-organelle lipidomics, enabling high-resolution lipid profiling in live cells. This advancement overcomes limitations of conventional methods, allowing detailed analysis of lipid heterogeneity within subcellular structures.
Area of Science:
- Biochemistry
- Cell Biology
- Spectroscopy
Background:
- Lipidomics is crucial for understanding cellular functions and diseases.
- Conventional lipidomics methods (e.g., mass spectrometry) require lipid extraction, preventing live-cell and single-organelle analysis.
- Existing techniques average data, masking vital cell-to-cell variations.
Purpose of the Study:
- To develop a high-resolution method for lipid profiling in live cells at the single-organelle level.
- To overcome the limitations of conventional lipidomics techniques.
- To enable quantitative analysis of organellar lipid heterogeneity.
Main Methods:
- Development of a micro-Raman assay for single-organelle lipidomics.
- Utilizing Raman microscopy for biochemical composition and lipidome analysis (concentration, unsaturation, isomer ratio, sphingolipids, cholesterol).
- Employing newly developed biomolecular component analysis software for data processing.
Main Results:
- Demonstrated sub-micrometer resolution lipid profiling in live cells, sufficient for subcellular structures.
- Successfully measured lipid concentration, unsaturation levels, cis/trans isomer ratios, sphingolipids, and cholesterol in single organelles.
- Established a robust protocol for quantitative lipid profile analysis in subcellular structures.
Conclusions:
- The micro-Raman assay enables single-organelle lipidomics in live or fixed cells.
- This method provides unprecedented insight into organellar lipid heterogeneity.
- Opens new quantitative avenues for studying cellular phenotypic variability in health and disease.
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