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