A pioneering technique of the Prevedal Group, EMBL allows neuroscientists to study live neurons deep inside the brain or other cells concealed within an opaque tissue. Elaborately, the technique is based on two microscopy methods: Three-photon microscopy and adaptive optics.
The details of the development is published in a paper in the journal Nature Methods.
In fact, before the technique was developed it was challenging for neuroscientists to study astrocytes that discharge calcium waves in deep layers of the cortex, or to imagine any other neural cells in the hippocampus. Importantly, the phenomenon occurs in the brains of all live mammals regularly.
Meanwhile, with the development of the technique, the research group were able to harness the fine details of the versatile cells at very high resolution.
In the field of neurosciences, brain tissues are mostly studied in small model organisms or samples of ex vivo that need to be sliced to be observed. This is because both of which represent non-physiological conditions.
Anatomically, normal brain cell functioning occurs only in live animals, however, the brain of the mouse is a highly scattering tissue. Due to such an anatomy, focus of light does not happen easily as it interacts with cellular components. This puts a limitation on how deep an image can be generated thus making it difficult to focus on small structures deep within the brain using traditional techniques.
On the other hand, using traditional fluorescence brain microscopy techniques, each time, two photons are absorbed by fluorescence molecule to make sure that the excitement due to the radiation is restricted to a small volume.
With further travel of the photons, the probability of being lost due to scattering increases. Increasing the wavelength of exciting photon toward infrared is one way to overcome the challenge.