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For GSDIM (Ground State Depletion with Individual Molecule return) microscopy using the Leica SR GSD microscope, Active Motif’s fluorescent Chromeo™ 546 dye and conjugated secondary antibodies have been tested by Leica Microsystems. The fluorescent properties of Chromeo 546 dye and secondaries meet the specifications required to perform GSDIM microscopy with the SR GSD system, using the 532 nm/500 mW laser for excitation. To ensure that you achieve the best image quality possible, Chromeo 546-stained slides should be embedded in a PVA (Polyvinyl alcohol) or Mowiol-based embedding media.
Figure 1: Chromeo 546 antibody conjugates in widefield and GSDIM microscopy.
Tubulin was stained with a primary monoclonal mouse antibody and with Chromeo 546 Goat anti-mouse IgG (Catalog No. 15033) secondary antibody. The widefield (left) and GSDIM (right) images are courtesy of Leica Microsystems, Germany.
Chromeo 546 is available as a reactive NHS-Ester or, for convenience in labeling your primary antibodies and proteins, offered in complete, optimized Fluorescent Antibody Labeling Kits.
