SUBCELLULAR VISUALIZATION OF LIGHT MICROSCOPIC SPECIMENS BY LASER SCANNING MICROSCOPY AND COMPUTER-ANALYSIS - A NEW APPLICATION OF IMAGE-ANALYSIS

被引:30
作者
ITOH, J [1 ]
OSAMURA, RY [1 ]
WATANABE, K [1 ]
机构
[1] TOKAI UNIV,SCH MED,DEPT PATHOL,ISEHARA,KANAGAWA 25911,JAPAN
关键词
LASER SCANNING MICROSCOPE; COMPUTER IMAGE ANALYSIS; LIVING CELLS; IMMUNOHISTOCHEMISTRY; ENZYME HISTOCHEMISTRY; SUBCELLULAR ORGANELLES;
D O I
10.1177/40.7.1607644
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
To identify subcellular organelles or to observe their pathological changes in sections prepared for light microscopy, immuno- and/or enzyme histochemical staining for the marker substances or enzymes of those subcellular organelles are frequently employed. With conventional light microscopes (CLM), however, it is hardly possible to determine whether or not the target organelles are properly stained and to confirm their fine structure. In the present study, the laser scanning microscope (LSM) was employed to obtain highly contrasted images of histochemically stained subcellular organelles at the limit of resolution in light microscopy. To refine or characterize those images, images built up as electronic signals in LSM were further processed in the Image Analysis System (IAS) with pipeline. Thus, the approximate figures of subcellular organelles such as microtubules, endoplasmic reticula, secretory granules, and mitochondria were visualized in brightfield on sections prepared for light microscopy (paraffin, frozen sections and cultured living cells). The validity of the images obtained by LSM or LSM-IAS was confirmed by immunoelectron microscopy when possible. The LSM images of histochemically stained suborganelles of various cells were definitely improved (refined and/or strengthened) by processing them with IAS.
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页码:955 / 967
页数:13
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