Medical Research Council - London Institute of Medical Sciences

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Multicolour three-photon fluorescence imaging deep into living mouse brain using single-wavelength excitation

11 April 2021

Deep Thinking

Peering deep into biological specimens is often a challenge for microscopists – light bends or diffracts as it hits thick tissues, and laser beams fired at fluorescently-stained structures can leave images blurred as out-of-focus light creeps in. Here a form of three-photon microscopy works around these problems, firstly by only 'switching on' fluorescent molecules energised by three photons at the same time, while a particular type of laser light helps to penetrate deep into this chunk of a mouse’s brain. Zooming down through the brain, researchers use a particular wavelength of laser that allows them to 'see' multiple colours at once – revealing blood vessels (red) mingling with neurons (green) and a mixture of blood cells and the brain’s myelin (blue). The techniques could be put to use in many different tissues, helping in the diagnosis of injury and disease.

Written by John Ankers

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