Orapim Tulyathan

Publications of Orapim Tulyathan

• Optical lock-in detection imaging microscopy for contrast-enhanced imaging in living cells.

  Authors: Gerard Marriott, Shu Mao, Tomoyo Sakata, Jing Ran, David K Jackson, Chutima Petchprayoon, Timothy J Gomez, Erica Warp, Orapim Tulyathan, Holly L Aaron, Ehud Y Isacoff, Yuling Yan

  Proceedings of the National Academy of Sciences of the United States of America. 12/2008; 105(46):17789-94.

  One of the limitations on imaging fluorescent proteins within living cells is that they are usually present in small numbers and need to be detected over a large background. We have developed theOne of the limitations on imaging fluorescent proteins within living cells is that they are usually present in small numbers and need to be detected over a large background. We have developed the means to isolate specific fluorescence signals from background by using lock-in detection of the modulated fluorescence of a class of optical probe termed "optical switches." This optical lock-in detection (OLID) approach involves modulating the fluorescence emission of the probe through deterministic, optical control of its fluorescent and nonfluorescent states, and subsequently applying a lock-in detection method to isolate the modulated signal of interest from nonmodulated background signals. Cross-correlation analysis provides a measure of correlation between the total fluorescence emission within single pixels of an image detected over several cycles of optical switching and a reference waveform detected within the same image over the same switching cycles. This approach to imaging provides a means to selectively detect the emission from optical switch probes among a larger population of conventional fluorescent probes and is compatible with conventional microscopes. OLID using nitrospirobenzopyran-based probes and the genetically encoded Dronpa fluorescent protein are shown to generate high-contrast images of specific structures and proteins in labeled cells in cultured and explanted neurons and in live Xenopus embryos and zebrafish larvae.

• Remote control of neuronal activity with a light-gated glutamate receptor.

  Authors: Stephanie Szobota, Pau Gorostiza, Filippo Del Bene, Claire Wyart, Doris L Fortin, Kathleen D Kolstad, Orapim Tulyathan, Matthew Volgraf, Rika Numano, Holly L Aaron, Ethan K Scott, Richard H Kramer, John Flannery, Herwig Baier, Dirk Trauner, Ehud Y Isacoff

  Neuron. 06/2007; 54(4):535-45.

  The ability to stimulate select neurons in isolated tissue and in living animals is important for investigating their role in circuits and behavior. We show that the engineered light-gated ionotropicThe ability to stimulate select neurons in isolated tissue and in living animals is important for investigating their role in circuits and behavior. We show that the engineered light-gated ionotropic glutamate receptor (LiGluR), when introduced into neurons, enables remote control of their activity. Trains of action potentials are optimally evoked and extinguished by 380 nm and 500 nm light, respectively, while intermediate wavelengths provide graded control over the amplitude of depolarization. Light pulses of 1-5 ms in duration at approximately 380 nm trigger precisely timed action potentials and EPSP-like responses or can evoke sustained depolarizations that persist for minutes in the dark until extinguished by a short pulse of approximately 500 nm light. When introduced into sensory neurons in zebrafish larvae, activation of LiGluR reversibly blocks the escape response to touch. Our studies show that LiGluR provides robust control over neuronal activity, enabling the dissection and manipulation of neural circuitry in vivo.

• Molecular imaging of hydrogen peroxide produced for cell signaling.

  Authors: Evan W Miller, Orapim Tulyathan, Orapim Tulyanthan, Ehud Y Isacoff, Christopher J Chang

  Nature chemical biology. 05/2007; 3(5):263-7.

  Hydrogen peroxide (H2O2) is emerging as a newly recognized messenger in cellular signal transduction. However, a substantial challenge in elucidating its diverse roles in complex biologicalHydrogen peroxide (H2O2) is emerging as a newly recognized messenger in cellular signal transduction. However, a substantial challenge in elucidating its diverse roles in complex biological environments is the lack of methods for probing this reactive oxygen metabolite in living systems with molecular specificity. Here we report the synthesis and application of Peroxy Green 1 (PG1) and Peroxy Crimson 1 (PC1), two new fluorescent probes that show high selectivity for H2O2 and are capable of visualizing endogenous H2O2 produced in living cells by growth factor stimulation, including the first direct imaging of peroxide produced for brain cell signaling. The combined features of reactive oxygen species selectivity, sensitivity to signaling levels of H2O2, and live-cell compatibility presage many new opportunities for PG1, PC1 and related synthetic reagents for exploring the physiological roles of H2O2 in living systems with molecular imaging.