Boslett Brianna, Nag Subhra, Resnick Andrew
Department of Biology, Geology, and Environmental Science, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, USA.
Department of Biology, Geology, and Environmental Science, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, USA ; Department of Physics, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, USA ; Center for Gene Regulation in Health and Disease, Cleveland State University, 2121 Euclid Avenue, Cleveland, OH 44115, USA.
Biomed Res Int. 2014;2014:532105. doi: 10.1155/2014/532105. Epub 2014 Mar 20.
Mycoplasma contamination of cultured cell lines is difficult to detect by routine observation. Infected cells can display normal morphology and the slow growth rate of mycoplasma can delay detection for extended periods of time, compromising experimental results. Positive identification of mycoplasma typically requires cells to be either fixed and stained for DNA or processed with PCR. We present a method to detect mycoplasma using live-cell optical microscopy typically used for routine observation of cell cultures. Images of untreated mycoplasma-infected epithelial cells alongside images of infected cells treated with Plasmocin, a commercially available antibiotic targeted to mycoplasma, are shown. We found that optical imaging is an effective screening tool for detection of mycoplasma contamination. Importantly, we found that cells regained normal function after the contamination was cleared. In conclusion, we present a technique to diagnose probable mycoplasma infections in live cultures without fixation, resulting in faster response times and decreased loss of cell material.
通过常规观察很难检测培养细胞系中的支原体污染。受感染的细胞可能呈现正常形态，而且支原体生长缓慢，可能会长时间延迟检测，从而影响实验结果。支原体的阳性鉴定通常需要对细胞进行固定、DNA染色或用聚合酶链反应（PCR）处理。我们提出了一种使用活细胞光学显微镜检测支原体的方法，这种显微镜通常用于细胞培养的常规观察。文中展示了未经处理的支原体感染上皮细胞的图像以及用Plasmocin（一种针对支原体的市售抗生素）处理的感染细胞的图像。我们发现光学成像对于检测支原体污染是一种有效的筛选工具。重要的是，我们发现污染清除后细胞恢复了正常功能。总之，我们提出了一种在不固定的情况下诊断活培养物中可能存在的支原体感染的技术，从而实现更快的响应时间并减少细胞材料的损失。
