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Protein Dynamics in Living Cells

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Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
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Updated: Sep 10, 2025

Proton Transfer and Protein Conformation Dynamics in Photosensitive Proteins by Time-resolved Step-scan Fourier-transform Infrared Spectroscopy

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Wavelength Dependence of Intact Protein Extraction Using Femtosecond Laser Ablation.

Alexander A C Wainwright¹, Khaled Madhoun¹, Pei Su^2,3,4,5

¹Dept. of Physics, University of Toronto, Toronto, ON Canada, M5R 2M8.

The Journal of Physical Chemistry Letters

|August 20, 2025

Summary

Femtosecond laser ablation enables intact protein extraction by carefully selecting laser wavelengths. Minimizing multiphoton ionization preserves protein signals for mass spectrometry imaging.

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Area of Science:

Analytical Chemistry
Biophysics
Spectroscopy

Background:

Femtosecond laser ablation (FLA) is a technique used for analyzing biological samples.
Intact protein extraction is crucial for high-resolution mass spectrometry imaging.
Wavelength-dependent ionization processes can affect protein integrity during FLA.

Purpose of the Study:

To investigate the influence of laser wavelength on intact protein extraction using FLA.
To determine optimal laser wavelengths for preserving protein signals.
To understand laser-biomolecule interactions in the context of mass spectrometry.

Main Methods:

Proteins were sampled using femtosecond laser wavelengths from ultraviolet to near-infrared.
Intact protein mixtures (9-68 kDa) and individual proteins were analyzed.
Wavelength-dependent multiphoton and avalanche ionization processes were studied.

Main Results:

Visible and infrared laser wavelengths facilitate intact protein extraction.
Ultraviolet wavelengths can also yield intact extraction if resonant absorption is avoided.
Minimizing multiphoton ionization is key to preserving intact protein signals.

Conclusions:

Laser wavelength selection is critical for successful intact protein extraction via FLA.
Understanding these interactions enhances the application of FLA in mass spectrometry imaging.
This research provides insights into high-intensity laser-biomolecule interactions.