Fig. 3: GelMap facilitates sample navigation for correlative live and expansion microscopy.

a, Schematic of the workflow for correlative live and expansion microscopy. (i) Cells of interest cultured on GelMap coverslips are followed live to image a dynamic process. (ii) Cells are incorporated into the ExM hydrogel, digested and the region of interest as indicated by the pattern is excised before expansion. (iii) Gel fragment is expanded and pattern is used to find back the cell of interest for correlation. b, Dissociated neurons cultured on GelMap navigation grid, stained for MAP2 (green), Tau (magenta), 4,6-diamidino-2-phenylindole (DAPI) (gray) and myc-tag (orange). c, Timelapse imaging of a mitotic cell stably expressing YFP–H2B and mCherry–tubulin. During anaphase, the cell was fixed on the microscope stage. The cell was located on navigation grids and expanded using TREx. d, Maximum projection of expanded cell from timelapse imaging in c stained for total protein using maleimide and DAPI (expansion factor, 10.7). Zooms are indicated by boxed regions. General protein stain reveals several ultrastructural features, including cell morphology, intracellular organelles such as mitochondria, the spindle midzone (yellow arrow), presumptive kinetochores (zoom 1, orange arrows) and centrioles (zoom 2). Linescan and quantification of centriole width indicated in orange and magenta. FWHM, average full width at half maximum for both linescans; AU, arbitrary units. Scale bars, 200 µm (b); timelapse 10 µm (c), below, 200 µm (left) and 100 µm (right); 5 µm (d), zoomed regions 1 µm. Scale bars in expanded samples reflect pre-expansion sizes.