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In order to facilitate versatile applications with high field dynamic nuclear polarization (DNP), it is important to be able to optimize the DNP performance, i.e. reach high nuclear hyperpolarization within a short signal build up time. Given that the solid-state DNP process is strongly temperature-dependent, it is important to benchmark the temperature dependence of various DNP and electron paramagnetic resonance (EPR) parameters that can then be used to test and develop theories and models for high field DNP mechanisms. However, DNP and EPR experiments at high fields and cryogenic temperatures below 20 Kelvin usually require home built instrumentation, and therefore even basic experimental observations are lacking in the literature. DNP and EPR experiments at 7 T (197 GHz) and 8.5 T (240 GHz), respectively, were conducted at temperatures between 35 K and 3.7 K where the electron thermal polarization changes from 13.4% to 85.6%, respectively. The samples are frozen solutions of 15 mM OX063Me trityl radicals in various mixtures of [1-13C]pyruvic acid, glycerol, and Gd3+-chelates. For all sample mixtures, the trityl EPR lines are found to be inhomogeneously broadened and the dominant DNP mechanism is shown to be the cross effect (CE). A 20%, 11%, and 6.77% 13C polarization is achieved at 3.7 K with a [1-13C]pyruvic–glycerol–H2O sample, the addition of 2 mM of Gd3+-chelates, and pure [1-13C]pyruvic acid, respectively. When T1n is sufficiently long, our results seem to suggest T1e is a key variable in the DNP process, where longer T1e values correlate with larger DNP enhancements (εDNP). The experimental data reported here on the temperature dependence of T1n, T1e, Tm (electron phase memory time), the EPR linewidth, TDNP and εDNP at high fields will be helpful for testing the mechanism and theory of DNP processes. [ABSTRACT FROM AUTHOR]

Interspin distances between 0.8 nm and 2.0 nm can be measured through the dipolar broadening of the continuous wave (cw) EPR spectrum of nitroxide spin labels at X-band (9.4 GHz, 0.35 T). We introduce Gd3+ as a promising alternative spin label for distance measurements by cw EPR above 7 Tesla, where the ☋−1/2〉 to ☋1/2〉 transition narrows below 1 mT and becomes extremely sensitive to dipolar broadening. To estimate the distance limits of cw EPR with Gd3+, we have measured spectra of frozen solutions of GdCl3 at 8.6 T (240 GHz) and 10 K at concentrations ranging from 50 mM to 0.1 mM, covering a range of average interspin distances. These experiments show substantial dipolar broadening at distances where line broadening cannot be observed with nitroxides at X-band. This data, and its agreement with calculated dipolar-broadened lineshapes, show Gd3+ to be sensitive to distances as long as ∼3.8 nm. Further, the linewidth of a bis-Gd3+ complex with a flexible ∼1.6 nm bridge is strongly broadened as compared to the mono-Gd3+ complex, demonstrating the potential for application to pairwise distances. Gd-DOTA-based chelates that can be functionalized to protein surfaces display linewidths narrower than aqueous GdCl3, implying they should be even more sensitive to dipolar broadening. Therefore, we suggest that the combination of tailored Gd3+ labels and high magnetic fields can extend the longest interspin distances measurable by cw EPR from 2.0 nm to 3.8 nm. cw EPR data at 260 K demonstrate that the line broadening remains clear out to similar average interspin distances, offering Gd3+ probes as promising distance rulers at temperatures higher than possible with conventional pulsed EPR distance measurements. [ABSTRACT FROM AUTHOR]