Kevin Claytor

Direct and cost-efficient hyperpolarization of long-lived nuclear spin states on universal 15N2-diazirine molecular tags

More than 10,000-fold enhanced magnetic resonance signals with >20-min signal lifetimes on universal biomolecular markers. Conventional magnetic resonance (MR) faces serious sensitivity limitations which can be overcome by hyperpolarization methods, but the most common method (dynamic nuclear polarization) is complex and expensive, and applications are limited by short spin lifetimes (typically seconds) of biologically relevant molecules. We use a recently developed method, SABRE-SHEATH, to directly hyperpolarize 15N2 magnetization and long-lived 15N2 singlet spin order, with signal decay time constants of 5.8 and 23 minutes, respectively. We find >10,000-fold enhancements generating detectable nuclear MR signals that last for over an hour. 15N2-diazirines represent a class of particularly promising and versatile molecular tags, and can be incorporated into a wide range of biomolecules without significantly altering molecular function.

Measuring long-lived 13C2 state lifetimes at natural abundance.

Long-lived disconnected eigenstates (for example, the singlet state in a system with two nearly equivalent carbons, or the singlet-singlet state in a system with two chemically equivalent carbons and two chemically equivalent hydrogens) hold the potential to drastically extend the lifetime of hyperpolarization in molecular tracers for in vivo magnetic resonance imaging (MRI). However, a first-principles calculation of the expected lifetime (and thus selection of potential imaging agents) is made very difficult because of the large variety of relevant intra- and intermolecular relaxation mechanisms. As a result, all previous measurements relied on costly and time consuming syntheses of (13)C labeled compounds. Here we show that it is possible to determine (13)C singlet state lifetimes by detecting the naturally abundant doubly-labeled species. This approach allows for rapid and low cost screening of potential molecular biomarkers bearing long-lived states.

Accessing Long-Lived Disconnected Spin-1/2 Eigenstates through Spins > 1/2

Pairs of chemically equivalent (or nearly equivalent) spin-1/2 nuclei have been shown to create disconnected eigenstates that are very long-lived compared with the lifetime of pure magnetization (T1). Here the classes of molecules known to have accessible long-lived states are extended to include those with chemically equivalent spin-1/2 nuclei accessed by coupling to nuclei with spin > 1/2, in this case deuterium. At first, this appears surprising because the quadrupolar interactions present in nuclei with spin > 1/2 are known to cause fast relaxation. Yet it is shown that scalar couplings between deuterium and carbon can guide population into and out of long-lived states, i.e., those immune from the dominant relaxation mechanisms. This implies that it may be practical to consider compounds with (13)C pairs directly bound to deuterium (or even (14)N) as candidates for storage of polarization. In addition, experiments show that simple deuteration of molecules with (13)C pairs at their natural abundance is sufficient for successful lifetime measurements.

Multicontrast nonlinear optical microscopy with a compact and rapid pulse shaper.

Homodyne detection can dramatically enhance measurement sensitivity for weak signals. In nonlinear optical microscopy it can make accessible a range of novel, intrinsic, contrast like nonlinear absorption and nonlinear phase contrast. Here a compact and rapid pulse shaper is developed, implemented, and demonstrated for homodyne detection in nonlinear microscopy with high-repetition rate mode-locked femtosecond lasers. With this method we generate two-photon absorption (TPA) and self-phase modulation images of gold nanostars in biological samples. Simultaneous imaging of two-photon luminescence and TPA also enables us to produce two-photon quantum yield images.

Accessing long lived (1)H states via (2)H couplings.

In this paper we demonstrate long-lived states involving a pair of chemically equivalent protons, with lifetimes ∼30 times T1 up to a total lifetime of ∼117s at high field (8.45T). This is demonstrated on trans-ethylene-d2 in solution, where magnetic inequivalence gives access to the long-lived states. It is shown that the remaining J-coupling between the two quadrupolar deuterium spins, JQQ, splits the conditions for optimally generating proton singlet states. Detailed simulations of the spin evolution are performed, shedding light on the coherent evolution during singlet-triplet conversion as well as on the incoherent evolution that causes relaxation. Subsequently, the simulations are compared with experimental results validating the theoretical insights. Possible applications include storage of hyperpolarization in the proton long-lived state. Of particular interest may be utilization of parahydrogen induced polarization to directly induce the examined long-lived states.

Nonlinear Pump-Probe Techniques for Multi-Contrast Microscopy

We demonstrate combined nonlinear transient absorption and nonlinear phase contrast imaging using rapid femtosecond pulse shaping techniques. We will discuss the implementation and potential applications of this technique for imaging biological specimens.

Pump-probe nonlinear phase dispersion spectroscopy: molecular contrast of pigmented and non-pigmented samples

Pump-probe nonlinear phase dispersion spectroscopy is a novel method that delivers molecular information of pigmented and non-pigmented samples by probing four dimensions: phase, amplitude, wavelength, and pump-probe time-delay. Its potential for molecular imaging is explored.

Femtosecond Pulse Shaping Enables Nonlinear Imaging in Highly Scattering Materials

Spectral re-shaping of pulses from a mode-locked femtosecond laser allows detection of two photon absorption and self phase modulation in highly scattering materials, which permits nonlinear tissue imaging with this intrinsic contrast.

Accessing nonlinear phase contrast in biological tissue using femtosecond laser pulse shaping

Nonlinear imaging takes advantage of the localized nature of the interaction to achieve high spatial resolution, optical sectioning, and deeper penetration in tissue. However, nonlinear contrast (other than fluorescence or harmonic generation) is generally difficult to measure because it is overwhelmed by the large background of detected illumination light. Especially challenging to measure is the nonlinear refractive index - accessing this quantity would allow the extension of widely employed phase microscopy methods to the nonlinear regime. We have developed a technique to suppress the background in these types of measurements by using femtosecond pulse shaping to encode nonlinear interactions in background-free regions of the frequency spectrum. Using this individual pulse shaping based technique we have been able to measure self-phase modulation (SPM) in highly scattering environments, such as biological tissue, with very modest power levels. Using our measurement technique we have demonstrated strong intrinsic SPM signatures of glutamate-induced neuronal activity in hippocampal brain slices. We have also extended this measurement method to cross-phase modulation, the two-color analogue to SPM. The two-color approach dramatically improves the measurement sensitivity by reducing undesired background and associated noise. We will describe the nonlinear phase contrast measurement technique and report on its application for imaging neuronal activity.