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Live single cell analysis using synchrotron FTIR microspectroscopy: development of a simple dynamic flow system for

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This study presents a cost-effective method for analyzing live biological cells using synchrotron radiation Fourier transform infrared microspectroscopy (SR-microFTIR). The modified system enables long-term cell viability and captures detailed biochemical changes, advancing live cell imaging.

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Area of Science:

  • Biophysics
  • Cell Biology
  • Spectroscopy

Background:

  • Synchrotron radiation Fourier transform infrared microspectroscopy (SR-microFTIR) offers detailed biochemical and morphological analysis of cells.
  • Analyzing live cells with SR-microFTIR is challenging due to water's infrared absorbance and limited cell viability outside culture conditions.

Purpose of the Study:

  • To develop a cost-effective method for live cell SR-microFTIR analysis.
  • To demonstrate extended cell viability and simultaneous measurement of biochemical changes in live cells.

Main Methods:

  • Simple, cost-effective modifications were made to a commercial liquid sample holder for SR-microFTIR.
  • Cell viability was assessed over 24 hours.
  • Live cells were analyzed at increasing temperatures to observe spectral changes.
  • Deuterated palmitic acid (D31-PA) uptake was studied over approximately 4 hours.

Main Results:

  • The modified system demonstrated cell viability for at least 24 hours.
  • Spectral changes in protein bands (α-β transition) were observed with increasing temperature, correlating with biochemical changes.
  • Over 200 spectra were collected for D31-PA uptake studies, showing the system's capability for extended measurements.
  • High spectral quality was maintained throughout the extended measurement periods.

Conclusions:

  • The developed SR-microFTIR system effectively supports live cell analysis with extended viability.
  • The system can simultaneously induce and measure biochemical changes in real-time.
  • Future developments aim to expand experimental capabilities, including drug-cell interaction studies.