使用激光捕获显微切割进行单细胞采样:一种从异质细胞群中收获靶细胞的技术

激光捕获显微切割有助于从载玻片上制备的细胞混合物中获取单个靶细胞。首先取一个合适的载玻片，在其表面涂有透紫外线、生化惰性膜。将载玻片暴露在紫外线下，使其更具亲水性，从而获得更好的细胞粘附性。

载玻片组装到细胞离心装置中。将细胞悬液添加到设备的漏斗端口上。细胞离心机，使离心力集中并将单层细胞沉积在载玻片上的小区域上。让载玻片风干。

将载玻片（样品面朝上）放在激光显微解剖显微镜下。定义感兴趣的单元格周围的区域。现在，聚焦激光束以解剖含有靶细胞的区域以及膜涂层。接下来，使激光束散焦并脉冲以产生压力。该能量将显微解剖区域弹射到预组装的收集管的倒盖中，该收集管包含放置在载玻片上方的缓冲液。

取出管子并盖上其盖子。旋转试管，将含有单个靶细胞的缓冲液带到试管底部。冷藏试管直至进一步分析。

对于激光微粒解剖，将整个 300 微升细胞悬液以 300 x g 的浓度离心到膜包被的载玻片上 5 分钟。然后，将膜涂层载玻片放在能够进行激光微镜解剖的显微镜上。接下来，将 4.5 微升细胞裂解预混液移入 0.2 毫升 PCR 管的盖子中。

将盖子放在样品上方，并通过激光显微切割收获单个细胞。此后，立即检查 PCR 帽中是否有分离的细胞。从激光显微解剖显微镜中取出PCR管并关闭管。短时间旋转收集管底部的所有液体。

• Microdissection - The process of extracting a single cell from a mixed cell population.
• Cytocentrifuge Technique - A method which uses centrifugal force to focus cells onto a slide.
• Target Capture - The retrieval of a specific designated cell from a slide.
• Laser Capture Microdissection Microscope - An instrument purposed to select and extract a target cell from a tissue section.
• Cytocentrifugation Meaning - The connotation of concentrating and depositing cells on a slide using centrifugal force.

• The concept of microdissection entails selecting and isolating a specific cell from a mix (e.g., target capture).
• Cytocentrifuge technique is a process employing centrifugal force to deposit cells on a slide (e.g., cytocentrifugation).
• Laser capture microdissection microscope allows for precise selection and extraction of target cells (e.g., microdissection).
• In context with the experiment, these principles are implemented to isolate a single target cell on a slide for further analysis.

• What does microdissection entail and what role does target capture play in it?
• From the provided information, can you explain the cytocentrifuge technique?
• What is the use of a laser capture microdissection microscope in the process?

• Microdissection has practical applications in cell study and genome research (e.g., biotechnology).
• These techniques contribute to the clinical and research aspects of healthcare industry (e.g., pathology).
• Isolation of specific cells has broader implications for disease understanding and treatment (e.g., cancer research).
• Advancements in efficient cell isolation techniques drive scientific innovation in cell and genome studies.