Schütze K, Pösl H, Lahr G
First Medical Department, Academic Hospital München-Harlaching, I. Med. Abteilung, München, Germany.
Cell Mol Biol (Noisy-le-grand). 1998 Jul;44(5):735-46.
The UV-laser microbeam has been established as a valuable tool in a wide area of molecular biology as well as in medical research and applications. This system allows to cut or fuse microscopically small specimen. An important application of the cutting laser is laser microbeam microdissection (LMM) combined with laser pressure catapulting (LPC), which allows to procure single cells or small homogeneous cell areas for subsequent molecular analysis in an entirely "non-contact" manner. With LMM minute tissue areas, single cells or chromosomes are microdissected and separated from their surroundings. Subsequently, LPC ejects the dissectates directly into the cap of a sample tube without any mechanical contact. This enables the rapid procurement of homogeneous specimen from less than one up to several hundreds of micrometers in diameter without encroachment of the adjacent region. The mRNA information of the selected specimen as well as of the remaining probe are well preserved, as demonstrated with laser isolated samples from a routinely prepared tissue section of a differentiated colorectal adenocarcinoma. Reverse transcription of specific mRNA coding for cytoplasmic beta-actin and subsequent hemi-nested PCR amplification was not impaired. Any kind of tissue, as well as single cells from different sources and even subcellular structures can be captured using this laser method. Wherever homogeneous samples are required to analyze cell or chromosome-specific genetic alterations such as in cancer research or prenatal diagnosis this unique and rapid laser micropreparation method will become a key technology of great value.
紫外激光微束已成为分子生物学广泛领域以及医学研究与应用中的一种重要工具。该系统能够对微观尺度的小样本进行切割或融合。切割激光的一项重要应用是结合激光压力弹射（LPC）的激光微束显微切割（LMM），它能以完全“非接触”的方式获取单个细胞或小的均匀细胞区域，用于后续的分子分析。通过LMM可对微小组织区域、单个细胞或染色体进行显微切割，并使其与周围环境分离。随后，LPC可将切割物直接弹射到样品管的管帽中，而无需任何机械接触。这使得能够从直径小于1微米到数百微米的范围内快速获取均匀样本，且不会侵犯相邻区域。所选样本以及剩余探针的mRNA信息能得到很好的保存，这在从分化型结肠腺癌常规制备的组织切片中通过激光分离的样本得到了证明。编码细胞质β-肌动蛋白的特定mRNA的逆转录以及随后的半巢式PCR扩增均未受到影响。使用这种激光方法可以捕获任何类型的组织，以及来自不同来源的单个细胞甚至亚细胞结构。在癌症研究或产前诊断等需要均匀样本以分析细胞或染色体特异性基因改变的任何情况下，这种独特且快速的激光微制备方法将成为一项极具价值的关键技术。
