Carey K. Johnson

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Two-photon scanning fluorescence microscopy has become a powerful tool for imaging living cells and tissues. Most applications of two-photon microscopy employ a Ti:sapphire laser excitation source, which is not readily portable or rapidly tunable. This work explores the use of two-photon fiber laser excitation (TP-FLEX) as an excitation source for scanning(More)
A voltage-controlled birefringent cell based on ceramic PMN-PT material is used to enable fast intensity modulation of femtosecond laser pulses in the 800 nm wavelength window. The birefringent cell based on a PMN-PT compound has comparatively high electro-optic response, allowing for a short interaction length of 3 mm and thus very small size, low(More)
Fluorescence correlation spectroscopy (FCS) is a robust method for the detection of intramolecular dynamics in proteins but is also susceptible to interference from other dynamic processes such as triplet kinetics and photobleaching. We describe an approach for the detection of intramolecular dynamics in proteins labeled with a FRET dye pair based on global(More)
We demonstrate a wavelength tunable optical excitation source for coherent Raman scattering (CRS) spectroscopy based on a single femtosecond fiber laser. Electrically controlled wavelength tuning of Stokes optical pulses was achieved with soliton self frequency shift in an optical fiber, and linear frequency chirping was applied to both the pump and the(More)
Fluorescence correlation spectroscopy (FCS) can be coupled with Förster resonance energy transfer (FRET) to detect intramolecular dynamics of proteins on the microsecond time scale. Here we describe application of FRET-FCS to detect fluctuations within the N-terminal and C-terminal domains of the Ca(2+)-signaling protein calmodulin. Intramolecular(More)
We demonstrate coherent Raman spectroscopy (CRS) using a tunable excitation source based on a single femto-second fiber laser. The frequency difference between the pump and the Stokes pulses was generated by soliton self-frequency shifting in a nonlinear optical fiber. Spectra of C─H stretches of cyclohexane were measured simultaneously by stimulated Raman(More)
We used single-pair fluorescence resonance energy transfer (spFRET) to track distance changes between domains of fluorescently labeled calmodulin (CaM) on the sub-millisecond time scale. In most cases, CaM remained in the same conformational substate over time periods of up to 1 ms, showing that conformational interchange occurs on a longer time scale.(More)
We demonstrate multi-mode microscopy based on a single femtosecond fiber laser. Coherent anti-Stokes Raman scattering (CARS), stimulated Raman scattering (SRS) and photothermal images can be obtained simultaneously with this simplified setup. Distributions of lipid and hemoglobin in sliced mouse brain samples and blood cells are imaged. The dependency of(More)
We analyze single molecule FRET burst measurements using Bayesian nested sampling. The MultiNest algorithm produces accurate FRET efficiency distributions from single-molecule data. FRET efficiency distributions recovered by MultiNest and classic maximum entropy are compared for simulated data and for calmodulin labeled at residues 44 and 117. MultiNest(More)
The speed and accuracy of Förster resonance energy transfer (FRET) measurements can be improved by rapidly alternating excitation wavelengths between the donor and acceptor fluorophore. We demonstrate FRET efficiency measurements based on a fiber laser and photonic crystal fiber as the source for two-photon excitation (TPE). This system offers the potential(More)