Abstract
We have been developing a 3-dimensional internal structure microscope for the observation of internal structures of samples. The internal structure is obtained by processing over 400 stored images of sliced faces of the sample, sliced continuously by the precise rotary slicer proceeding under observation with a microscope. 3D-ISM has been utilized to observe food, farm products and biological organs. Under white light, it is difficult to get clear images of expressed gene under high magnification. In order to overcome the problem, we installed a fluorescence microscope function into the 3D-ISM for detection of fluorescent materials. The performance of the apparatus are as follows. The resolution of the sample sending is 0.5μm. The maximum observation speed is five sections per second. The lowest sample cooling temperature is -50°C. The object lens magnification are from 2 to 100. U and B and G excitations are available by the fluorescent wave length. Each sliced face image is recorded on a postscript type laser disk after sliced, followed the next slicing. With the recorded section's image a 3-dimensional image is constructed on a workstation by volume rendering method, and hence the internal of the sample is observable. A cancer field gene (c-kit) in a embryo of ICR strain mice which is 11.5 day olds was observed. The embryo was dyed with a FITC labeled DNA probe (c-kit) using whole mount fluorescence in situ hybridization method. The existence of fluorescence inside the embryo's vertebra and internal organs were confirmed, and the topography of c-kit was observable. A device which is possible in fluorescent observation was constructed on trial, and succeeded in observation of topography of the gene in an embryo, which was difficult so far.
