Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry and Czech Advanced Technology and Research Institute, Palacký University Olomouc, Hněvotínská 5, 779 00 Olomouc, Czech Republic.
Int J Mol Sci. 2021 Jul 1;22(13):7098. doi: 10.3390/ijms22137098.
Cytocentrifugation is a common technique for the capture of cells on microscopic slides. It usually requires a special cytocentrifuge or cytorotor and cassettes. In the study presented here, we tested the new concept of cytocentrifugation based on the threaded connection of the lid and the sample holder to ensure an adjustable flow of solutions through the filters and the collection of the filtered solutions in the reservoir during centrifugation. To test this concept, we developed a device for the preparation of cell samples on circular coverslips. The device was tested for the capture and sample processing of both eukaryotic and prokaryotic cells, cell nuclei, and mitochondria for microscopy analysis including image cytometry. Moreover, an efficient procedure was developed for capturing formaldehyde-fixed cells on non-treated coverslips without cell drying. The results showed that the tested arrangement enables the effective capture and processing of all of the tested samples and the developed device represents an inexpensive alternative to common cytocentrifuges, as only the paper filter is consumed during sample processing, and no special centrifuge, cytorotor, or cassette is necessary. As no additional system of solution removal is required during sample staining, the tested concept also facilitates the eventual automation of the staining procedure.
细胞离心沉淀是一种将细胞捕获在显微镜载玻片上的常用技术。它通常需要特殊的细胞离心机或细胞旋转器和试剂盒。在本研究中,我们测试了基于盖子和样品架的螺纹连接来确保溶液通过过滤器的可调流量以及在离心过程中在储液器中收集过滤溶液的新概念。为了测试这一概念,我们开发了一种用于在圆形盖玻片上制备细胞样品的设备。该设备用于捕获和处理真核细胞和原核细胞、细胞核和线粒体,用于显微镜分析,包括图像细胞术。此外,还开发了一种用于在未经处理的盖玻片上捕获甲醛固定细胞而不干燥细胞的有效方法。结果表明,所测试的布置能够有效地捕获和处理所有测试的样品,所开发的设备代表了一种廉价的替代普通细胞离心机的方案,因为在样品处理过程中仅消耗滤纸,而不需要特殊的离心机、细胞旋转器或试剂盒。由于在样品染色过程中不需要额外的溶液去除系统,因此所测试的概念也便于最终实现染色过程的自动化。