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Fluorogenic Probes for Multicolor Imaging in Living Cells

Kai Johnsson, Luc Reymond, Grazvydas Lukinavicius, Olivier Vincent Sallin, Keitaro Umezawa

Here we present a far-red, silicon-rhodamine-based fluorophore (SiR700) for live-cell multicolor imaging. SiR700 has excitation and emission maxima at 690 nm and 715 nm, respectively. SiR700-based probes for F-actin, microtubules, lysosomes and SNAP-tag are fluorogenic, cell-permeable and compatible with superresolution microscopy. In conjunction with probes based on the previously introduced carboxy-SiR650, SiR700-based probes permit multicolor live-cell superresolution microscopy in the far-red, thus significantly expanding our capacity for imaging living cells.

American Chemical Society2016

SiR–Hoechst is a far-red DNA stain for live-cell nanoscopy

Kai Johnsson, Luc Reymond, Alberto Schena, Grazvydas Lukinavicius

Cell-permeable DNA stains are popular markers in live-cell imaging. Currently used DNA stains for live-cell imaging are either toxic, require illumination with blue light or are not compatible with super-resolution microscopy, thereby limiting their utility. Here we describe a far-red DNA stain, SiR–Hoechst, which displays minimal toxicity, is applicable in different cell types and tissues, and is compatible with super-resolution microscopy. The combination of these properties makes this probe a powerful tool for live-cell imaging.

2015

Fluorescent Labeling of SNAP-Tagged Proteins in Cells

Kai Johnsson, Luc Reymond, Grazvydas Lukinavicius

One of the most prominent self-labeling tags is SNAP-tag. It is an in vitro evolution product of the human DNA repair protein O6 -alkylguanine-DNA alkyltransferase (hAGT) that reacts specifically with benzylguanine (BG) and benzylchloropyrimidine (CP) derivatives, leading to covalent labeling of SNAP-tag with a synthetic probe (Gronemeyer et al., Protein Eng Des Sel 19:309–316, 2006; Curr Opin Biotechnol 16:453–458, 2005; Keppler et al., Nat Biotechnol 21:86–89, 2003; Proc Natl Acad Sci U S A 101:9955– 9959, 2004). SNAP-tag is well suited for the analysis and quantification of fused target protein using fluorescence microscopy techniques. It provides a simple, robust, and versatile approach to the imaging of fusion proteins under a wide range of experimental conditions. © Springer Science+Business Media New York 2015.

Springer2015