Preparation, Characterization, and Application of Optical Switch Probes
Published Online: 1 AUG 2010
Copyright © 2010 by John Wiley and Sons, Inc.
Lab Protocol Title
Current Protocols in Chemical Biology
How to Cite
Petchprayoon, C. and Marriott, G. 2010. Preparation, Characterization, and Application of Optical Switch Probes. Current Protocols in Chemical Biology. 2:153–169.
- Published Online: 1 AUG 2010
- Published Print: AUG 2010
Optical switches represent a new class of molecular probe with applications in high-contrast imaging and optical manipulation of protein interactions. Small molecule, organic optical switches based on nitrospirobenzopyran (NitroBIPS) and their reactive derivatives and conjugates undergo efficient, rapid and reversible, orthogonal optically-driven transitions between a colorless spiro (SP)-state and a colored merocyanine (MC)-state. The excited MC-state also emits fluorescence, which serves as the readout of the state of the switch. Defined optical perturbations of SP and MC generate a defined waveform of MC-fluorescence that can be isolated against unmodulated background signals by using a digital optical lock-in detection approach or to control specific dipolar interactions on proteins. The protocols describe general procedures for the synthesis and spectroscopic characterization of NitroBIPS and specifically labeled conjugates along with methods for the manipulation of dipolar interactions on proteins and imaging of the MC-state of NitroBIPS within living cells. Curr. Protoc. Chem. Biol. 2:153-169 © 2010 by John Wiley & Sons, Inc.
- optical switch;