Department of Physiology and Pathophysiology, University of Veterinary and Pharmaceutical Sciences, Brno, Czech Republic.
Arch Oral Biol. 2010 Aug;55(8):570-5. doi: 10.1016/j.archoralbio.2010.05.007. Epub 2010 Jun 15.
Laser capture microdissection (LCM) uniquely allows the selection of specific cell populations from histological sections. These selected cells are then catapulted into a test tube without any contamination from surrounding tissues. During the last ten years, many significant results have been achieved, particularly at the level of DNA and RNA where amplification techniques are available. However, where amplification procedures are difficult, the benefits of LCM diminish. To overcome such difficulties, a novel approach, combining laser capture microdissection and flow cytometry, has been tested here for detection of apoptosis and proliferation in tissue bound cell populations without any amplification steps. The mouse cap stage molar tooth germ was used as a model. At the centre of the inner enamel epithelium, the primary enamel knot is a clearly defined apoptotic population with minimal proliferation, flanked by the highly proliferative cervical loops on each side. Thus within the tooth germ epithelium at this stage, two distinct populations of cells are found side by side. These populations were selected by laser capture microdissection and then analysed by flow cytometry for apoptosis and proliferation. Flow cytometric results correlated well with immunohistochemical findings, demonstrating the success and sensitivity of this combined procedure.
激光捕获显微切割（LCM）独特地允许从组织切片中选择特定的细胞群体。然后，这些选定的细胞被弹射到一个试管中，不会受到周围组织的任何污染。在过去的十年中，已经取得了许多重大成果，特别是在 DNA 和 RNA 水平上，因为有扩增技术可用。然而，在扩增程序困难的情况下，LCM 的益处会减少。为了克服这些困难，这里测试了一种新的方法，将激光捕获显微切割和流式细胞术结合起来，用于检测组织结合细胞群体中的细胞凋亡和增殖，而无需任何扩增步骤。以小鼠帽状期磨牙牙胚为模型。在内釉上皮的中心，初级釉结是一个明确界定的凋亡群体，增殖很少，两侧为高度增殖的颈环。因此，在这个阶段的牙胚上皮中，发现了两个相邻的不同细胞群体。这些群体通过激光捕获显微切割进行选择，然后通过流式细胞术分析细胞凋亡和增殖。流式细胞术结果与免疫组织化学结果很好地相关，证明了这种联合程序的成功和敏感性。
