Magnetic resonance imaging using a nonuniform Bo (NuBo) field-cycling magnet.

Magnetic resonance imaging (MRI) is a powerful noninvasive diagnostic tool with superior soft tissue contrast. However, access to MRI is limited since current systems depend on homogeneous, high field strength main magnets (B0-fields), with strong switchable gradients which are expensive to install and maintain. In this work we propose a new approach to MRI where imaging is performed in an inhomogeneous field using radiofrequency spatial encoding, thereby eliminating the need for uniform B0-fields and conventional cylindrical gradient coils. The proposed technology uses an innovative data acquisition and reconstruction approach by integrating developments in field cycling, parallel imaging and non-Fourier based algebraic reconstruction. The scanner uses field cycling to image in an inhomogeneous B0-field; in this way magnetization is maximized during the high field polarization phase, and B0 inhomogeneity effects are minimized by using a low field during image acquisition. In addition to presenting the concept, this work provides experimental verification of a long-lived spin echo signal, spatially varying resolution, as well as both simulated and experimental 2D images. Our initial design creates an open MR system that can be installed in a patient examination table for body imaging (e.g., breast or liver) or built into a wall for weighted-spine imaging. The proposed system introduces a new class of inexpensive, open, silent MRIs that could be housed in doctor's offices much like ultrasound is today, making MRI more widely accessible.

Practical method for RF pulse distortion compensation using multiple square pulses for low-field MRI.

Since recovery time of the RF coil is long at low field MRI, the rising and the ring-down times of the square pulse are also long, which means the applied sinc pulse can easily be distorted from the changing amplitude. However, both the rising time and ring-down time can be calculated using Q-factor. Using this information, an RF square pulse were compensated by appending two square pulses before and after the RF pulse. The durations of these RF square pulses were calculated using the Q-factor. Since the amplitude of the sinc pulse changes continuously, a series of square pulses were applied to apply sinc pulse to the coil. The minimum number of square pulses and the amplitude of the square pulses were calculated. It was successfully demonstrated that the sinc pulse can be compensated using a series of square pulses. The more number of square pulses were used, the smoother sinc pulse was applied to the RF coil. The Q-factor was experimentally calculated from the ring-down time of a signal induced in a sniffer loop which was connected to an oscilloscope. The resulting Q-factor was then used to calculate both the duration and amplitude of the square pulses for compensation. Echo trains were also acquired in an inhomogeneous B0 field using the compensated RF pulses. In order to enhance the SNR of the echo trains, a pre-polarization pulse was added to the CPMG spin echo sequence. The SNRs of the echo signal acquired using compensated pulses were compared with those of signal obtained with uncompensated pulses and showed significant improvements of 61.1% and 51.5% for the square and sinc shaped pulses respectively.

Optimal Detection of Fusion Pore Dynamics Using Polarized Total Internal Reflection Fluorescence Microscopy.

The fusion pore is the initial narrow connection that forms between fusing membranes. During vesicular release of hormones or neurotransmitters, the nanometer-sized fusion pore may open-close repeatedly (flicker) before resealing or dilating irreversibly, leading to kiss-and-run or full-fusion events, respectively. Pore dynamics govern vesicle cargo release and the mode of vesicle recycling, but the mechanisms are poorly understood. This is partly due to a lack of reconstituted assays that combine single-pore sensitivity and high time resolution. Total internal reflection fluorescence (TIRF) microscopy offers unique advantages for characterizing single membrane fusion events, but signals depend on effects that are difficult to disentangle, including the polarization of the excitation electric field, vesicle size, photobleaching, orientation of the excitation dipoles of the fluorophores with respect to the membrane, and the evanescent field depth. Commercial TIRF microscopes do not allow control of excitation polarization, further complicating analysis. To overcome these challenges, we built a polarization-controlled total internal reflection fluorescence (pTIRF) microscope and monitored fusion of proteoliposomes with planar lipid bilayers with single molecule sensitivity and ∼15 ms temporal resolution. Using pTIRF microscopy, we detected docking and fusion of fluorescently labeled small unilamellar vesicles, reconstituted with exocytotic/neuronal v-SNARE proteins (vSUVs), with a supported bilayer containing the cognate t-SNAREs (tSBL). By varying the excitation polarization angle, we were able to identify a dye-dependent optimal polarization at which the fluorescence increase upon fusion was maximal, facilitating event detection and analysis of lipid transfer kinetics. An improved algorithm allowed us to estimate the size of the fusing vSUV and the fusion pore openness (the fraction of time the pore is open) for every event. For most events, lipid transfer was much slower than expected for diffusion through an open pore, suggesting that fusion pore flickering limits lipid release. We find a weak correlation between fusion pore openness and vesicle area. The approach can be used to study mechanisms governing fusion pore dynamics in a wide range of membrane fusion processes.