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Dual-Color Fluorescence Cross-Correlation Spectroscopy to Study Protein-Protein Interaction and Protein Dynamics in Live Cells

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Rapid-scan coherent 2D fluorescence spectroscopy.

Simon Draeger, Sebastian Roeding, Tobias Brixner

|March 1, 2017

Summary

This summary is machine-generated.

We developed a new method for coherent two-dimensional electronic spectroscopy in liquids using fluorescence detection. This technique enables rapid acquisition of 2D spectra, improving efficiency for studying molecular dynamics.

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Area of Science:

Physical Chemistry
Spectroscopy
Chemical Physics

Background:

Coherent two-dimensional electronic spectroscopy (2D ES) is a powerful technique for probing ultrafast molecular dynamics.
Traditional 2D ES methods can be time-consuming due to the need for scanning time delays.
Fluorescence detection offers high sensitivity but extracting coherent signals can be challenging.

Purpose of the Study:

To develop a rapid and efficient pulse-shaper-assisted coherent 2D electronic spectroscopy technique for liquid samples.
To demonstrate the capability of extracting coherent information from incoherent fluorescence signals.
To analyze the performance and signal-to-noise ratio of the developed method.

Main Methods:

Development of a customized pulse shaper for shot-to-shot modulation at 1 kHz.
Implementation of rapid scanning over all time delays for accelerated data acquisition.
Utilization of 27-step phase cycling to extract coherent signals from fluorescence.
Application of the technique to cresyl violet in ethanol.

Main Results:

Acquisition of a full 15 × 15 pixel 2D spectrum in approximately 6 seconds.
Successful recovery of known molecular oscillations as a function of population time.
Demonstration of a root-mean-square error < 0.05 after 1 minute of measurement time with rapid scanning.
Analysis of signal-to-noise ratio as a function of averaging.

Conclusions:

Pulse-shaper-assisted coherent 2D electronic spectroscopy with fluorescence detection is a viable and efficient technique for liquid samples.
The method allows for rapid acquisition of high-quality 2D electronic spectra.
This advancement facilitates faster and more detailed investigations of molecular dynamics in condensed phases.