Soitu Cristian, Panea Mirela, Castrejón-Pita Alfonso A, Cook Peter R, Walsh Edmond J
Osney Thermofluids Institute, Department of Engineering Science, University of Oxford, Osney Mead, Oxford OX2 0ES, United Kingdom.
Neurosciences Group, Nuffield Department of Clinical Neurosciences, Weatherall Institute of Molecular Medicine, John Radcliffe Hospital, Oxford OX3 9DS, United Kingdom.
Biomicrofluidics. 2021 Feb 8;15(1):014108. doi: 10.1063/5.0043312. eCollection 2021 Jan.
Many wound-healing assays are used in cell biology and biomedicine; they are often labor intensive and/or require specialized and costly equipment. We describe a contactless method to create wounds with any imaginable 2D pattern in cell monolayers using the micro-jets of either media or an immiscible and biocompatible fluorocarbon (i.e., FC40). We also combine this with another method that allows automation and multiplexing using standard Petri dishes. A dish is filled with a thin film of media overlaid with FC40, and the two liquids are reshaped into an array of microchambers within minutes. Each chamber in such a grid is isolated from others by the fluid walls of FC40. Cells are now added, allowed to grow into a monolayer, and wounds are created using the microjets; then, healing is monitored by microscopy. As arrays of chambers can be made using media and Petri dishes familiar to biologists, and as dishes fit seamlessly into their incubators, microscopes, and workflows, we anticipate that this assay will find wide application in wound healing.
细胞生物学和生物医学中使用了许多伤口愈合检测方法;这些方法通常劳动强度大,并且/或者需要专门且昂贵的设备。我们描述了一种非接触式方法,可使用培养基或不混溶且具有生物相容性的氟碳化合物(即FC40)的微喷射在细胞单层中创建具有任何可想象的二维图案的伤口。我们还将此方法与另一种方法相结合,该方法允许使用标准培养皿实现自动化和多重检测。将一个培养皿装满覆盖有FC40的薄培养基膜,两种液体在几分钟内重新形成微腔阵列。在这样的网格中,每个腔室通过FC40的流体壁与其他腔室隔离。现在添加细胞,使其生长成单层,然后使用微喷射创建伤口;然后,通过显微镜监测愈合情况。由于可以使用生物学家熟悉的培养基和培养皿制作腔室阵列,并且由于培养皿可以无缝地放入他们的培养箱、显微镜和工作流程中,我们预计这种检测方法将在伤口愈合中得到广泛应用。