Grötz K A, Piepkorn B, Bittinger F, Reichert T E, Duschner H, de Aguiar E G, Kann P, Wagner W
Universitätsklinik für Mund-, Kiefer- und Gesichtschirurgie, Johannes Gutenberg Universität Mainz, Germany.
Mund Kiefer Gesichtschir. 1998 May;2(3):141-5. doi: 10.1007/s100060050049.
Fixation (formalin), decalcification (sections) or mechanical treatment (grinding) all bear the risk of artifacts occurring during hard-tissue histology. Because studies on the etiology of pathological changes mostly focus on subclinical lesions, artifacts can simulate early changes or even be superimposed on existing changes. The objective of this study was to determine how artifacts can be reduced.
In confocal laser scanning microscopy (CLSM) a focused laser beam scans the surface of the specimens and penetrates into the tissue. The intensity of the remitted light is recorded. The confocal effect is due to an extremely small aperture (pin-hole), excluding light from out-of-focus planes of the sample. By stepwise movement of the object table, a tomographic series of tomographic images is obtained. Sound cortical bone samples of the lower jaw (n = 20) were studied by light microscopy and by CLSM, visualizing identical areas of a ground sectioned sample after H&E staining. Additionally, embedded and fresh blocks of tissue of the same bone sample were studied histotomographically in the CLSM.
(1) Light microscopic micromorphology of cortical bone can be visualized adequately in the CLSM; (2) many structures that can be visualized by light microscopy only after special staining (e.g., osteozyte processes) can be visualized by the CLSM using sample blocks without pretreatment.
(1) Nondestructive subsurface histotomography by CLSM totally excludes mechanical artifacts; (2) physicochemical artifacts can be handled more easily because fresh samples can be studied; (3) pseudo-three-dimensional imaging allows histological interpretation of the tissue that is equivalent to macroscopic tomographic techniques (CT, MRT).
固定（福尔马林）、脱钙（切片）或机械处理（研磨）在硬组织组织学检查过程中均存在产生伪像的风险。由于对病理变化病因的研究大多集中在亚临床病变上，伪像可能会模拟早期变化，甚至叠加在已有的变化之上。本研究的目的是确定如何减少伪像。
在共聚焦激光扫描显微镜（CLSM）中，聚焦激光束扫描标本表面并穿透组织。记录反射光的强度。共聚焦效应归因于一个极小的孔径（针孔），可排除来自样品非聚焦平面的光线。通过载物台的逐步移动，可获得一系列断层图像。对下颌骨的健康皮质骨样本（n = 20）进行光学显微镜和CLSM研究，在苏木精-伊红（H&E）染色后观察磨片样本的相同区域。此外，还在CLSM中对同一骨样本的包埋组织块和新鲜组织块进行了组织断层扫描研究。
（1）CLSM能够充分显示皮质骨的光学显微镜微观形态；（2）许多仅在特殊染色后才能通过光学显微镜观察到的结构（如骨细胞突起），使用未经预处理的样本块在CLSM中即可观察到。
（1）CLSM进行的无损表面下组织断层扫描完全排除了机械伪像；（2）由于可以研究新鲜样本，物理化学伪像更易于处理；（3）伪三维成像能够对组织进行组织学解释，这与宏观断层扫描技术（CT、MRT）相当。
