Department of Pathology and Cell Biology and Montreal Diabetes Center, University of Montreal, Canada.
J Histochem Cytochem. 2011 Oct;59(10):899-907. doi: 10.1369/0022155411418507. Epub 2011 Aug 10.
Low-voltage (5-kV) transmission electron microscopy revealed a novel aspect of the pancreatic acinar cell secretory granules not previously detected by conventional (80-kV) transmission electron microscopy. Examination of ultra-thin (30-nm) sections of non-osmicated, stain-free pancreatic tissue sections by low-voltage electron microscopy revealed the existence of granules with non-homogeneous matrix and sub-compartments having circular or oval profiles of different electron densities and sizes. Such partition is completely masked when observing tissues after postfixation with osmium tetroxide by low-voltage transmission electron microscopy at 5 kV and/or when thicker sections (70 nm) are examined at 80 kV. This morphological partition reflects an internal compartmentalization of the granule content that was previously predicted by morphological, physiological, and biochemical means. It corresponds to the segregation of the different secretory proteins inside the granule as demonstrated by high-resolution immunocytochemistry and reflects a well-organized aggregation of the secretory proteins at the time of granule formation in the trans-Golgi. Such partition of the granule matrix undergoes changes under experimental conditions known to alter the secretory process such as stimulation of secretion or diabetes.
低电压(5kV)透射电子显微镜揭示了胰腺腺泡细胞分泌颗粒的一个新方面,这是以前用常规(80kV)透射电子显微镜未检测到的。通过低电压电子显微镜检查未经锇固定、无染色的胰腺组织超薄(30nm)切片,发现存在颗粒基质不均匀,并且具有不同电子密度和大小的圆形或椭圆形轮廓的亚区室。当用 5kV 的低电压透射电子显微镜观察用四氧化锇后固定的组织时,或者当在 80kV 下检查更厚的切片(70nm)时,这种分区完全被掩盖。这种形态分区反映了颗粒内容物的内部分隔化,这是以前通过形态学、生理学和生物化学手段预测的。它对应于在颗粒形成时,不同分泌蛋白在颗粒内的分离,这如通过高分辨率免疫细胞化学所证明的,并反映了在分泌蛋白的有组织聚集。这种颗粒基质的分区在已知改变分泌过程的实验条件下发生变化,例如刺激分泌或糖尿病。