Spontaneously blinking JFb dyes

Overview

Applying the tuning strategies inherent in the Janelia Fluor® platform to hydroxymethyl (HM) derivatives of rhodamines yields a palette of spontaneously blinking dyes for super-resolution imaging. These dyes switch between fluorescent and nonfluorescent forms due to transient protonation/deprotonation allowing super-resolution microscopy without the need for redox buffers or strong illumination.

Key Benefits

• Simplified single-molecule localization microscopy (SMLM) workflows: No need for strong redox buffers or prebleaching protocols, ensuring every molecule gets counted.
• A broad series of on:off ratios to accommodate different labeling strategies, labeling densities, cellular environments, and microscopy techniques: Users can choose the right dye for their experiment rather than trying to make a single dye work.
• Exceptional brightness and photostability: Built on the Janelia Fluor® scaffold ensuring high brightness and robust photostability.
• Useful beyond SMLM: Compatible with super-resolution optical fluctuation imaging (SOFI) in live and fixed cells.

Highlighted Molecules

• JF₅₂₆b: A bright, green-emitting dye for localization microscopy that spontaneous blinks in standard imaging buffers.
• JF₆₃₅b: An all-around far-red-excited label for SMLM and SOFI compatible with HaloTag and standard chemical labeling techniques.
• Other far-red blinking dyes: JFX₆₅₀b, JF₆₄₆b, JF₆₃₉b, JF₆₃₀b, JF₆₂₉b, JF₆₂₆b, and JF₆₁₄b.

Applications

• Single-molecule super-resolution (SMLM)
• Super-resolution optical fluctuation imaging (SOFI)

References: ACS Cent. Sci. 2019, 5, 1602–1613, https://doi.org/10.1021/acscentsci.9b00676; bioRxiv 2024, https://doi.org/10.1101/2024.05.12.593749.

IP Coverage: US Patent Application US17/027,286

Offer: Available for non-exclusive commercial license.

Janelia Tech ID: 2019-054