Journal: Cells

Article Title: Correlative Multi-Modal Microscopy: A Novel Pipeline for Optimizing Fluorescence Microscopy Resolutions in Biological Applications

doi: 10.3390/cells12030354

Figure Lengend Snippet: Correlative Confocal-dSTORM Analysis on Image-Cytometry re-localized cells. ( a ) Representative re-localized G1 cells acquired by the correlative Confocal-STORM protocol described in the text. Confocal Z-Stacks were first acquired of the Tubulin-Atto425 and γH2A.X-Alexa488 fluorescence channel, followed by acquisition of the 53BP1-Cy3 and DynLL1-Alexa Fluor647 channels in dSTORM. The dSTORM plane is identified in the Z-Stack, aligned and merged in the plane (merged image in the second row on the left) and then inserted into the Z-stack (3D projection on the right). ( b ) The described correlative microscopy procedure was applied to cells in the different stages of mitosis, isolated according to the image-cytometry analysis. Z-stacks merged with the dSTORM plane of some representative cells are shown. Scale bar: 5 µm.

Article Snippet: Cells were then rinsed 3 times in PBS and incubated for 1 h with donkey anti-Rat IgG (H + L) biotin (a18749, ThermoFisher Scientific, Waltham, MA, USA) and for 45 min at room temperature with streptavidin Atto425 conjugated (S000-51, Rockland Immunochemicals, Pottstown, PA, USA).

Techniques: Cytometry, Fluorescence, Microscopy, Isolation