Cryoplane fluorescence microscopy

There is an urgent need for rapid 3D imaging methods to characterize transgenic lines of experimental animals. The novel instrument described here, a cryoplane fluorescence microscope, meets that need. It offers the required speed, high spatial resolution, and support for imaging of reporter genes. The instrument is a large horizontal cryostat equipped with macro- and microimaging facilities. It accomodates both brightfield imaging of systemically delivered counter stains and fluorescence imaging of probes introduced either by genetic manipulation or by direct injection. Spatial distortion in the resulting 3D datasets is quite low, less than 2%. We demonstrate here application of the instrument for mapping the expression pattern of the gene reporter GFP and for anatomical characterization using the nonfluorescent crystal violet.

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