snap tag labeling  (New England Biolabs)

Journal: HardwareX

Article Title: The miEye: Bench-top super-resolution microscope with cost-effective equipment

doi: 10.1016/j.ohx.2022.e00368

Figure Lengend Snippet: dSTORM imaging of fluorescently labeled nuclear pore complexes (NPCs) of fixed U2OS-Nup96-SNAP cells. A) Averaged image of the unprocessed 20,000 frame-long image stack showing an individual nucleus of a U2OS cell. B) Reconstructed super-resolution image of Nup96-SNAP-AF647-containing NPCs located at the distinct positions of a single U2OS cell’s entire nucleus. C) Close-up view of a chosen nucleus area (indicated by a white box in panel B ). Individual NPCs and their ring-like arrangement are visible. D) Fourier ring correlation (FRC) curve indicating the obtained spatial resolution of 34.7 nm.

Article Snippet: After quenching the remaining FA, a few drops of Image-iT FX Signal Enhancer (I36933, Thermo Fisher Scientific) were added onto the sample and incubated for 30 min. Staining of SNAP-tag labeled Nup96 was carried out at room temperature for 2 h using benzylguanine-conjugated AF647 (S9136S, New England Biolabs) diluted to the final concentration of 1 µM in PBS supplemented with 1 µM of dithiothreitol (R0862, Thermo Fisher Scientific) and 0.5 % (w/v) bovine serum albumin (T844.2, Carl Roth).

Techniques: Imaging, Labeling