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Diagram of multicolor brain imaging with a microscope-coupled fiber-bundle microprobe: GM, galvanometrically scanned mirrors; Obj, objective; L1, L2, lenses; F1, F2, F3, filters; PMT, photomultipliers.
Typical images of neuron circuits in the brain of a Thy1-EGFP mouse obtained through a fiber bundle coupled to a confocal microscope by using the fluorescence response from EGFP with the distal end of the fiber probe immersed at different depth in a fixed brain sample: (a) a raw image delivered by the fiber, (b) an image of the same area after Gaussian filtering with σ = 2.5 μm, (c) a stack of images obtained with a fiber-bundle microprobe immersed at different depths inside a fixed brain tissue, and (d) a typical image of individual neurons in the brain tissue obtained with a standard confocal microscope. The scale bar in the images is 50 μm.
Two-color images of dentate gyrus in the hippocampus of a transgenic Thy-1-EGFP mice additionally stained with DAPI dilactate acquired in a living mouse with the use of the fiber-bundle microprobe (a) and with fixed brain slices from the same mouse by means of confocal microscopy (b). The scale bar is 100 μm (a) and 50 μm (b).
Three-color image of dentate gyrus in the hippocampus of a transgenic Thy-1-EGFP mice double-stained with DAPI dilactate and Neurotrace Nissl 640/660 dye obtained with the use of the fiber-bundle microprobe. The scale bar is 100 μm.
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