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Single Cell Multiplex Reverse Transcription Polymerase Chain Reaction After Patch-clamp
Published on: June 20, 2018
Combination of patch clamp and Raman spectroscopy for single-cell analysis
Ute Neugebauer1, Stefan H Heinemann, Michael Schmitt
1Institute of Photonic Technology, Albert-Einstein-Strasse 9, 07745 Jena, Germany.
Analytical Chemistry
|December 15, 2010
Summary
This study introduces a novel method combining patch clamp and Raman spectroscopy for label-free detection of intracellular components. The technique successfully tracked the internalization of compounds like crocin and heme into living cells.
Area of Science:
- Biophysics
- Cell Biology
- Analytical Chemistry
Background:
- Patch clamp provides controlled access to the cell cytosol.
- Raman spectroscopy offers label-free detection of molecular components.
- Quantitative intracellular analysis is crucial for understanding cellular processes.
Purpose of the Study:
- To develop and validate a combined patch clamp and Raman spectroscopy technique.
- To enable label-free, quantitative detection of intracellular substances.
- To investigate the internalization kinetics of specific compounds into living cells.
Main Methods:
- Utilizing patch clamp for controlled cellular access and compound internalization.
- Employing Raman spectroscopy for label-free, real-time monitoring of intracellular analytes.
- Applying the technique to study crocin and heme internalization in mammalian cells.
Main Results:
- Demonstrated proof-of-principle with the carotinoid crocin, characterizing its internalization kinetics.
- Successfully differentiated the internalization of free heme versus protein-bound heme (MP-11).
- Observed accumulation of free heme due to binding with intracellular components.
Conclusions:
- The combined patch clamp-Raman spectroscopy method is effective for label-free quantitative intracellular analysis.
- This technique allows for the study of compound internalization and cellular interactions in living cells.
- The findings provide insights into the differential uptake and fate of molecules within cells.
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