A New Attractive Approach of Three- Dimensional(3D) Reconstruction of the Relationships Between Hormone Secreating Cells and its Microvessel Network Patterns in Rat Pituitary Gland by Confocal Laser Scanning Miroscopy(CLSM) With Combined Fluorescent and Non-Fluorescent Probes.

Abstract

Abstract There have been considerable interests in the relationship between hormone secreting endocrine cells and microvessel network patterns in human pituitary gland. However, its 3D microcirculatory networks have not been studied extensively. Therefore, this study was designed to visualize hormone secreting endcrine cells and microvessels in 3D by CLSM and to reveal relationship between hormone secretion and microvessel networks in the rat pituitary gland. CLSM has been an exciting new apparatus because of its more powerful resolution than conventional light microscopy and the high-performance on 3D optical sectioning. The images taken with CLSM can be digitized and made susceptible to image analysis manipulation and computer-assisted 3D reconstructions. The female normal adult Wistar rats were used in this study. To demonstrate microvessels, fluorescent probe( 30 mg/ml)-conjugated gelatin solution was injected to left ventricle of the heart After complete perfusion, the pituitary glands were resected and immediately fixed in cold graded PFA(0.5- 8 %) containing 15 % picric acid solutions (4 °C. GH, ACTH cells or S100 protein positive fcfliculostellate (FS) cells were demonstrated by indirect TRTTC-labeled or HRP labeled antibody methods.