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High contrast CARS microscopy – imaging of membrane proteins with high spatial resolution

G. M. Arzumanyan, K. Z. Mamatkulov

A new promising and highly sensitive approach to the imaging of protein crystals based on polarization-sensitive coherent anti-Stokes light scattering (P-CARS) has been successfully developed at FLNP based on the CARS laser scanning microspectrometer with fast data acquisition and high contrast vibrational imaging.

It should be noted that serial crystallography on X-ray synchrotron radiation sources and on free electron lasers of the latest generation has allowed to carry out data acquisition of crystals of micrometer and even sub-micrometer sizes that has resulted in significant progress in structural biology. However, the imaging of small crystals, being highly demanded, is still a task to meet, especially for membrane protein crystals. CARS microscopy provides an advanced non-destructive and marker-free imaging technique with high sensitivity and high lateral spatial resolution that allows to obtain chemically selective images of the basic types of macromolecules: proteins, lipids, nucleic acids, others.

Bioimaging can bring together investigations from different areas of life sciences, demonstrating innovations in cell imaging methodologies and image analysis techniques, as well as their application methods in practice.

References:

  • M. Arzumanyan and others, JACS, 2016, DOI: 10.1021/jacs.6b04464

Fig. 1. Imaging of a bacteriorhodopsin crystal: (a) microphotograph, (b) spontaneous Raman and (c) polarization-sensitive CARS