New approach speeds up Brillouin microscopy
Engineers have improved the speed and throughput of Brillouin microscopy 1000-fold.
A novel approach for Brillouin microscopy, developed by engineers at the European Molecular Biology Laboratory (EMBL; Heidelberg, Germany), could transform how researchers study light-sensitive biological samples. This new approach significantly enhances the speed and efficiency of imaging, offering a quicker and more precise method to explore the mechanical properties of samples in 3D.
The mechanical properties of cells and tissues, such as elasticity and viscosity, play a crucial role in cell function. However, current techniques for determining the mechanical properties of cells have limitations, such as requiring physical contact, being limited to surfaces or having poor resolution.
Brillouin microscopy, first developed in the early 2000s, relies on the phenomenon of Brillouin scattering: a process where light interacts with the natural thermal vibrations within a material, shifting the frequency of the light. The spectrum of scattered light can be measured, revealing the physical characteristics of a material.
However, there are several challenges with Brillouin microscopy, one being slow image acquisition, with early iterations only allowing a single pixel to be viewed at a time. In 2022, developments in Brillouin microscopy allowed for an expanded field of view from one pixel to a line of 100 pixels. While this is a significant advancement, engineers were still on a quest to make imaging acquisition even quicker. Now, EMBL engineers have developed a novel approach that enables light-sensitive organisms to be viewed more efficiently.
Picture perfect: imaging the full 3D orientation and position of molecules in cells
This novel hybrid microscope allows for the simultaneous imaging of the full 3D orientation and position of molecules within cells.
“Over the years, we have progressed from being able to see just a pixel at a time to a line of 100 pixels, to now a full plane that offers a view of approximately 10,000 pixels,” commented lead author Carlo Bevilacqua.
The engineers were able to use the symmetric properties of the Brillouin spectrum and a custom-built Fourier-transform imaging spectrometer to demonstrate full-field 2D spectral Brillouin imaging of both phantoms and biological samples at a throughput of 40,000 spectra per second, with a precision of nearly 70MHz. Altogether, this method improved the speed and throughput in Brillouin microscopy 1000-fold, allowing researchers to capture 2D and 3D images of biological samples, with minimal light exposure, much faster than before.
“Just as the development of light-sheet microscopy here at EMBL marked a revolution in light microscopy because it allowed for faster, high-resolution, and minimally phototoxic imaging of biological samples, so too does this advance in the area of mechanical or Brillouin imaging,” commented corresponding author Robert Prevedel.
The EMBL engineers hope that this new approach will open more opportunities for researchers and the bioimaging community.
