How should I establish pre-migration/invasion reference wells?
如何建立移民前/入侵参考井？
Pre-migration/invasion reference wells are used to establish the size and position of the detection zone in order to quantify the extent of movement in experimental wells. For Oris assays, you can leave some stoppers in place until you read the results. Since Oris Pro assays lack stoppers, this method is not applicable. For Oris Pro and Oris assays, alternative methods to establish reference wells are: Simply collect images at time zero as migration is about to begin. Add fixative to replicate wells or a replicate plate at the beginning of the assay. However, when fixing cells in a subset of wells in the plate, this method risks impacting cells adjacent to fixed wells as they may be exposed to fixative vapors. Add an inhibitor of migration or invasion, such as cytochalasin D, to reference wells at the beginning of the assay. Be sure to add sufficient inhibitor to block cell movement completely. You may wish to read a related application note here.
迁移前/入侵参考井用于确定检测区的大小和位置，以便量化实验井中的移动范围。 对于 Oris 测定仪，您可以在读取结果之前保留一些塞子。由于 Oris Pro 测定仪没有塞子，因此此方法不适用。 对于 Oris Pro 和 Oris 检测，可采用其他方法建立参照井： 只需在迁移即将开始的零时收集图像即可。 在检测开始时将固定液加入复制孔或复制板中。不过，在固定平板中部分孔的细胞时，这种方法有可能会影响邻近固定孔的细胞，因为它们可能会暴露在固定液蒸汽中。 检测开始时，在参照孔中加入迁移或侵袭抑制剂，如细胞松弛素 D。确保加入足够的抑制剂以完全阻止细胞移动。 您可以阅读相关的应用说明 这里。