Morphometry in anatomy: image analysis on fine structure and histochemical reactions with special reference to radioautography.

Anatomy now has a step to transfer from qualitative science to quantitative science. The methods of anatomical investigation employ optical instruments. Anatomy initially described qualitative features of the structure of the body, but now it turns to transfer to new fields measuring the number, size, length, area and volume of not only, structure but also the histochemical components of the structure with quantitative analysis. In order to quantify histochemical reactions in various organs, light and electron microscopic radioautograms were prepared from the liver, pancreas, intestines and kidney which were labeled with 3H-thymidine, 3H-uridine, 3H-leucine, 3H-glucosamine and 3H-glycerol, fixed in glutaraldehyde and osmium tetroxide, embedded in Epoxy resin, sectioned either thick or thin, coated with either Konica NR-M2 or -H2 radioautographic emulsions, exposed, developed, stained for either light or electron microscopic radioautographs. The radioautographs were observed with Olympus Vanox AHB-LB or Nikon Optiphot-2 light microscopes or Hitachi H-700, JEOL JEM-4000EX electron microscopes at 200 and 400 kV. The radioautograms were analysed with various image and analysed such as PPA-250, MOP, Digigramer-G, IBAS-II, Quadra 900, Luzex 300 to count the number of silver grains and to measure the size and number of structures. Some electron microscopic radioautographs were analysed in analytical electron microscopes, combined with Hitachi H-700 Horiba EMAX-1800E, JEOL JEM-200CX Kevex 7000-77, and JEOL JEM-4000EX Tracor-Northern TN-5400 to quantify silver grain contents. The results demonstrated that correct data can be obtained by controlling both full-automatic and semi-automatic image analysers by operating them manually.