Unit

Preparation, Characterization, and Application of Optical Switch Probes

  1. Chutima Petchprayoon,
  2. Gerard Marriott

Published Online: 1 AUG 2010

DOI: 10.1002/9780470559277.ch100054

Current Protocols in Chemical Biology

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.

Author Information

  1. Department of Bioengineering, University of California, Berkeley, California

Publication History

  1. Published Online: 1 AUG 2010
  2. Published Print: AUG 2010

Abstract

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.

Keywords:

  • optical switch;
  • nitrospiropyran;
  • imaging