Figure 1
From: Heterogeneity of miR-10b expression in circulating tumor cells
Schematic representation of the combined ISH/IF procedure.
The three day stepwise protocol for analysis of CTCs detected with the CellSearch® system. CTC enrichment and staining initially occurs using the CellSearch®. CTCs are then enumerated using the CellTracks® Analyzer and fixed in the CellSearch® cartridge. The ISH/IHC protocol using LNA probes proceeds in cartridge, followed by manual visualization of miR expression in CTCs on a cell-by-cell basis. DIG labelled probes detected with FITC labelled anti-DIG antibody allowed visualisation and localization of miR expression in cells, which are also re-labelled with cell specific, CTC identifying antibodies from the Circulating Epithelial Cell Kit (Veridex). CTCs can then be labelled as either miR+, CK+, DAPI+ and CD45−; or miR−, CK+, DAPI+ and CD45−, in a semi-quantitative way.
