Obtaining High-Quality Hyperpolarized Xenon-129 Magnetic Resonance Images

<ol>
	<li>Pellegrino, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interpretative+strategies+for+lung+function+tests.">Interpretative strategies for lung function tests.</a> European Respiratory Journal. 26 (5), 948-968 (2005).</li><li>Ebner, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperpolarized+129Xenon+MRI+to+quantify+regional+ventilation+differences+in+mild+to+moderate+asthma:+A+prospective+comparison+between+semi-automated+ventilation+defect+percentage+calculation+and+pulmonary+function+tests.">Hyperpolarized 129Xenon MRI to quantify regional ventilation differences in mild to moderate asthma: A prospective comparison between semi-automated ventilation defect percentage calculation and pulmonary function tests.</a> Investigative Radiology. 52 (2), 120-127 (2017).</li><li>Abuelhia, E., Alghamdi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+arising+exposure+of+ionizing+radiation+from+computed+tomography+and+the+associated+health+concerns.">Evaluation of arising exposure of ionizing radiation from computed tomography and the associated health concerns.</a> Journal of Radiation Research and Applied Sciences. 13 (1), 295-300 (2020).</li><li>Kern, A. L., Vogel-Claussen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperpolarized+gas+MRI+in+pulmonology.">Hyperpolarized gas MRI in pulmonology.</a> The British Journal of Radiology. 91 (1084), 20170647 (2018).</li><li>M&#246;ller, H. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+of+the+lungs+using+hyperpolarized+noble+gases.">MRI of the lungs using hyperpolarized noble gases.</a> Magnetic Resonance in Medicine. 47 (6), 1029-1051 (2002).</li><li>Salerno, M., Altes, T. A., Mugler, J. P., Nakatsu, M., Hatabu, H., de Lange, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperpolarized+noble+gas+MR+imaging+of+the+lung:+Potential+clinical+applications.">Hyperpolarized noble gas MR imaging of the lung: Potential clinical applications.</a> European Journal of Radiology. 40 (1), 33-44 (2001).</li><li>. New Drug Therapy Approvals at 2022 Available from: <a target="_blank" href="https://www.fda.gov/drugs/new-drugs-fda-cders-new-molecular-entities-and-new-therapeutic-biological-products/new-drug-therapy-approvals-2022">https://www.fda.gov/drugs/new-drugs-fda-cders-new-molecular-entities-and-new-therapeutic-biological-products/new-drug-therapy-approvals-2022</a> (2023)</li><li>Nikolaou, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Near-unity+nuclear+polarization+with+an+open-source+129Xe+hyperpolarizer+for+NMR+and+MRI.">Near-unity nuclear polarization with an open-source 129Xe hyperpolarizer for NMR and MRI.</a> Proceedings of the National Academy of Sciences. 110 (35), 14150-14155 (2013).</li><li>Birchall, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=XeUS:+A+second-generation+automated+open-source+batch-mode+clinical-scale+hyperpolarizer.">XeUS: A second-generation automated open-source batch-mode clinical-scale hyperpolarizer.</a> Journal of Magnetic Resonance. 319, 106813 (2020).</li><li>He, M., Zha, W., Tan, F., Rankine, L., Fain, S., Driehuys, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+comparison+of+two+hyperpolarized+129Xe+MRI+ventilation+quantification+pipelines:+The+effect+of+signal+to+noise+ratio.">A comparison of two hyperpolarized 129Xe MRI ventilation quantification pipelines: The effect of signal to noise ratio.</a> Academic Radiology. 26 (7), 949-959 (2019).</li><li>Niedbalski, P. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protocols+for+multi-site+trials+using+hyperpolarized+129+Xe+MRI+for+imaging+of+ventilation,+alveolar-airspace+size,+and+gas+exchange:+A+position+paper+from+the+129+Xe+MRI+clinical+trials+consortium.">Protocols for multi-site trials using hyperpolarized 129 Xe MRI for imaging of ventilation, alveolar-airspace size, and gas exchange: A position paper from the 129 Xe MRI clinical trials consortium.</a> Magnetic Resonance in Medicine. 86 (6), 2966-2986 (2021).</li><li>M&#246;ller, H. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+of+the+lungs+using+hyperpolarized+noble+gases.">MRI of the lungs using hyperpolarized noble gases.</a> Magnetic Resonance in Medicine. 47 (6), 1029-1051 (2002).</li><li>Bier, E. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+thermally+polarized+129Xe+phantom+for+quality+assurance+in+multi-center+hyperpolarized+gas+MRI+studies.">A thermally polarized 129Xe phantom for quality assurance in multi-center hyperpolarized gas MRI studies.</a> Magnetic Resonance in Medicine. 82 (5), 1961-1968 (2019).</li><li>Wild, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+between+2D+and+3D+gradient-echo+sequences+for+MRI+of+human+lung+ventilation+with+hyperpolarized+3He.">Comparison between 2D and 3D gradient-echo sequences for MRI of human lung ventilation with hyperpolarized 3He.</a> Magnetic Resonance in Medicine. 52 (3), 673-678 (2004).</li><li>Garrison, W. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Lung+volume+dependence+and+repeatability+of+hyperpolarized+129Xe+MRI+gas+uptake+metrics+in+healthy+volunteers+and+participants+with+COPD.">Lung volume dependence and repeatability of hyperpolarized 129Xe MRI gas uptake metrics in healthy volunteers and participants with COPD.</a> Radiology: Cardiothoracic Imaging. 5 (3), e220096 (2023).</li><li>Ni, W., Qi, J., Liu, L., Li, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+pulse+signal+preprocessing+method+based+on+the+Chauvenet+criterion.">A pulse signal preprocessing method based on the Chauvenet criterion.</a> Computational and Mathematical Methods in Medicine. 2019, 2067196 (2019).</li><li>. Available from: <a target="_blank" href="https://www.129xectc.org">https://www.129xectc.org</a> (2023)</li></ol>

Robert Thomen has provided consulting to Polarean, LLC.

The ability to troubleshoot 129Xe MRI issues is a necessary skill and may help mitigate problems in real time. Until a hyperpolarized gas infrastructure can be purchased from a single party and garner support from scanner manufacturers, these quality control tasks are the sole responsibility of the individual laboratories. The goal of this manuscript is to provide the reader with helpful practices and suggestions for the inevitable event of poor data acquisition. While we attempt to address as many potential i...

For over a century, lung function assessment has primarily relied on global measurements from spirometry and body plethysmography. However, these traditional pulmonary function tests (PFTs) are limited in their ability to capture early-stage disease&#39;s regional nuances and subtle changes in lung tissue1. Nuclear medicine with inhaled radiotracers has been used widely for the assessment of ventilation/perfusion mismatches commonly associated with pulmonary emboli, but this involves ionizing radiation and yields lower resolution. In contrast, computed tomography (CT) has emerged as the gold standard for lung imaging, offering exceptional spatial and temporal clarity compared to nuclear imaging2. While low-dose CT scans can mitigate radiation exposure, potential radiation risk should still be considered3,4. Proton MRI of the lung is uncommon due to low tissue density of the lung and rapid signal decay from lung tissue, although recent advances offer functional information despite potential low signal. On the other hand, hyperpolarized xenon magnetic resonance imaging (HP 129Xe MRI) is a non-invasive modality that allows for imaging of lung function with regional specificity5,6. It produces a high nonequilibrium nuclear magnetization of the gas in liter quantities. The inert gas is then inhaled by a subject inside the MR scanner for a single breath and is directly imaged by the scanner. Thus, the inhaled gas is directly imaged as opposed to the tissue itself. This technique has been used to assess lung ventilation across many diseases, including asthma, chronic obstructive pulmonary disease (COPD), cystic fibrosis, idiopathic pulmonary fibrosis, coronavirus disease 2019 (COVID-19), and many others3. In December 2022, HP 129Xe MRI was approved by the United States FDA as an MRI ventilation contrast agent to be used in the United States of America (USA) in adults and pediatric patients aged 12 years and older7. Physicians can now use 129Xe MRI to better care for patients with improved/personalized treatment plans.
Historically, clinical MRI focuses exclusively on imaging hydrogen nuclei (protons) which are abundant in nearly all human viscera. The MRI scanners, sequences, and quality control are generally maintained by the scanner manufacturer as part of the site license and warranty. However, 129Xe requires a multinuclear capable MR scanner and has required a dedicated research team to operationalize the hyperpolarizer, custom-built radiofrequency (RF) coils, dedicated pulse sequences, and offline reconstruction/analysis software. Each of these components can be supplied by third-party vendors or developed in-house. Thus, the burden of quality control generally rests on the 129Xe research team as opposed to the scanner manufacturer or individual third party. Consistent acquisition of high-quality 129Xe data is therefore uniquely challenging as each component of the 129Xe MRI process introduces the potential for error, which must be closely monitored by the 129Xe team. Not only can these situations be extremely frustrating as researchers have to troubleshoot and investigate possible causes for any challenges that may have arisen, but they can be very costly as this slows down patient imaging and subject recruitment. Some costs associated with troubleshooting involve MRI time costs, the hyperpolarization of 129Xe, which involves the consumption of different gases, and the use of materials. Additionally, with the recent FDA approval and growth in 129Xe imaging, providing a standardized protocol for quality control is necessary to avoid common issues and setbacks in 129Xe operation8,9.
Here, we present some of the more commonly encountered issues in 129Xe MRI, including RF coil failures, the emergence of various noise profiles that lead to low signal-to-noise ratio (SNR), and poor quality images10. We aim to provide some concise quality control (QC) guidelines and protocols to ensure the acquisition of high-quality image data and troubleshoot some of the more common issues that can arise in 129Xe MRI. The insights provided here are also relevant for hyperpolarized helium-3 troubleshooting.

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	<tbody>
		<tr>
			<td>Polarization measurement station&#160;</td>
			<td>Polerean</td>
			<td>42881</td>
			<td>https://polarean.com/</td>
		</tr>
		<tr>
			<td>Pressure vessele with plunger valve</td>
			<td>Ace glass</td>
			<td>8648-85</td>
			<td>https://www.aceglass.com/html/3dissues/Pressure_Vessels/offline/download.pdf</td>
		</tr>
		<tr>
			<td>Tedlar bag</td>
			<td>Jensen inert</td>
			<td>GST381S-0707TJO&#160;&#160;</td>
			<td>http://www.jenseninert.com/</td>
		</tr>
		<tr>
			<td>Xenon Hyperpolarizer 9820</td>
			<td>Polerean</td>
			<td>49820</td>
			<td>https://polarean.com/</td>
		</tr>
		<tr>
			<td>Xenon loop coil</td>
			<td>Clinical MR Solutions</td>
			<td>Custom device</td>
			<td>https://www.sbir.gov/sbc/clinical-mr-solutions-llc</td>
		</tr>
		<tr>
			<td>Xenon vest coil</td>
			<td>Clinical MR Solutions</td>
			<td>Custom device</td>
			<td>https://www.sbir.gov/sbc/clinical-mr-solutions-llc</td>
		</tr>
	</tbody>
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troubleshooting and quality assurance in hyperpolarized xenon magnetic resonance imaging: tools for high-quality image acquisition

Hyperpolarized (HP) xenon magnetic resonance imaging (129Xe MRI) is a recently federal drug administration (FDA)-approved imaging modality that produces high-resolution images of an inhaled breath of xenon gas for investigation of lung function. However, implementing 129Xe MRI is uniquely challenging as it requires specialized hardware and equipment for hyperpolarization, procurement of xenon imaging coils and coil software, development and compilation of multinuclear MR imaging sequences, and reconstruction/analysis of acquired data. Without proper expertise, these tasks can be daunting, and failure to acquire high-quality images can be&#160;frustrating, and expensive. Here, we present some quality control (QC) protocols, troubleshooting practices, and helpful tools for129Xe MRI sites, which may aid in the acquisition of optimized, high-quality data and accurate results. The discussion will begin with an overview of the process for implementing HP 129Xe MRI, including requirements for a hyperpolarizer lab, the combination of 129Xe MRI coil hardware/software, data acquisition and sequence considerations, data structures, k-space and image properties, and measured signal and noise characteristics. Within each of these necessary steps lies opportunities for errors, challenges, and unfavorable occurrences leading to poor image quality or failed imaging, and this presentation aims to address some of the more commonly encountered issues. In particular, identification and characterization of anomalous noise patterns in acquired data are necessary to avoid image artifacts and low-quality images; examples will be given, and mitigation strategies will be discussed. We aim to make the 129Xe MRI implementation process easier for new sites while providing some guidelines and strategies for real-time troubleshooting.

Figure 4&#160;depicts the results of the noise characterization analysis performed on the noise scan. The plot demonstrates the impact of both regular and irregular noise on the k-space, where the deviation from the ideal y=x reference line is observed. Regular noise leads to a continuous pattern in the k-space, while irregular noise results in high-value outliers in the QQ plot.
Moving on to Figure 5, a series of lung images acquired...

Watch this Scientific Journal Video about Using Hyperpolarized Xenon-129 MRI to Study Lung Diseases at JoVE.com

Author Spotlight: Using Hyperpolarized Xenon-129 MRI to Study Lung Diseases 

Here, we present a protocol for obtaining high-quality hyperpolarized xenon-129 magnetic resonance images, covering hardware, software, data acquisition, sequence selection, data management, k-space utilization, and noise analysis.

Troubleshooting and Quality Assurance in Hyperpolarized Xenon Magnetic Resonance Imaging: Tools for High-Quality Image Acquisition

Hyperpolarized (HP) xenon magnetic resonance imaging (129Xe MRI) is a recently federal drug administration (FDA)-approved imaging modality that produces ...

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1.1K Views.  University of Missouri. Here, we present a protocol for obtaining high-quality hyperpolarized xenon-129 magnetic resonance images, covering hardware, software, data acquisition, sequence selection, data management, k-space utilization, and noise analysis.

Video: Author Spotlight: Using Hyperpolarized Xenon-129 MRI to Study Lung Diseases 

Visualizing Proteins and Macromolecular Complexes by Negative Stain EM: from Grid Preparation to Image Acquisition

Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural biology 1. In recent years, this method has achieved great success in studying structures of proteins and macromolecular complexes 2, 3. Compared with electron cryomicroscopy (cryoEM), in which frozen hydrated protein samples are embedded in a thin layer of vitreous ice 4, negative staining is a simpler sample preparation method in which protein samples are embedded in a thin layer of dried heavy metal salt to increase specimen contrast 5. The enhanced contrast of negative stain EM allows examination of relatively small biological samples. In addition to determining three-dimensional (3D) structure of purified proteins or protein complexes 6, this method can be used for much broader purposes. For example, negative stain EM can be easily used to visualize purified protein samples, obtaining information such as homogeneity/heterogeneity of the sample, formation of protein complexes or large assemblies, or simply to evaluate the quality of a protein preparation.
In this video article, we present a complete protocol for using an EM to observe negatively stained protein sample, from preparing carbon coated grids for negative stain EM to acquiring images of negatively stained sample in an electron microscope operated at 120kV accelerating voltage. These protocols have been used in our laboratory routinely and can be easily followed by novice users.

Visualizing protein samples by negative stain electron microscopy (EM) has become a popular structural analysis method. It is useful for quantitative structural analysis, such as calculating a 3D reconstruction of the molecules being studied, and also for qualitative examination of the quality of protein preparations. In this article we present detailed protocols for preparing the EM grids, staining the sample and visualizing the sample in an electron microscope. Novice users can follow these protocols easily and to utilize negative stain EM as a routine assay, in addition to other biochemical assays, for evaluating their protein samples.

Single particle electron microscopy (EM), of both negative stained or frozen hydrated biological samples, has become a versatile tool in structural ...

Magnetic Resonance Elastography Methodology for the Evaluation of Tissue Engineered Construct Growth

Traditional mechanical testing often results in the destruction of the sample, and in the case of long term tissue engineered construct studies, the use of destructive assessment is not acceptable. A proposed alternative is the use of an imaging process called magnetic resonance elastography. Elastography is a nondestructive method for determining the engineered outcome by measuring local mechanical property values (i.e., complex shear modulus), which are essential markers for identifying the structure and functionality of a tissue. As a noninvasive means for evaluation, the monitoring of engineered constructs with imaging modalities such as magnetic resonance imaging (MRI) has seen increasing interest in the past decade1. For example, the magnetic resonance (MR) techniques of diffusion and relaxometry have been able to characterize the changes in chemical and physical properties during engineered tissue development2. The method proposed in the following protocol uses microscopic magnetic resonance elastography (&mu;MRE) as a noninvasive MR based technique for measuring the mechanical properties of small soft tissues3. MRE is achieved by coupling a sonic mechanical actuator with the tissue of interest and recording the shear wave propagation with an MR scanner4. Recently, &mu;MRE has been applied in tissue engineering to acquire essential growth information that is traditionally measured using destructive mechanical macroscopic techniques5. In the following procedure, elastography is achieved through the imaging of engineered constructs with a modified Hahn spin-echo sequence coupled with a mechanical actuator. As shown in Figure 1, the modified sequence synchronizes image acquisition with the transmission of external shear waves; subsequently, the motion is sensitized through the use of oscillating bipolar pairs. Following collection of images with positive and negative motion sensitization, complex division of the data produce a shear wave image. Then, the image is assessed using an inversion algorithm to generate a shear stiffness map6. The resulting measurements at each voxel have been shown to strongly correlate (R2&gt;0.9914) with data collected using dynamic mechanical analysis7. In this study, elastography is integrated into the tissue development process for monitoring human mesenchymal stem cell (hMSC) differentiation into adipogenic and osteogenic constructs as shown in Figure 2.

The procedure demonstrates the methodology of magnetic resonance elastography for monitoring the engineered outcome of adipose and osteogenic tissue engineered constructs through noninvasive local assessment of the mechanical properties using microscopic magnetic resonance elastography (&mu;MRE).

Traditional mechanical testing often results in the destruction of the sample, and in the case of long term tissue engineered construct studies, the use ...

Magnetically-Assisted Remote Controlled Microcatheter Tip Deflection under Magnetic Resonance Imaging

X-ray fluoroscopy-guided endovascular procedures have several significant limitations, including difficult catheter navigation and use of ionizing radiation, which can potentially be overcome using a magnetically steerable catheter under MR guidance.
	The main goal of this work is to develop a microcatheter whose tip can be remotely controlled using the magnetic field of the MR scanner. This protocol aims to describe the procedures for applying current to the microcoil-tipped microcatheter to produce consistent and controllable deflections.
	A microcoil was fabricated using laser lathe lithography onto a polyimide-tipped endovascular catheter. In vitro testing was performed in a waterbath and vessel phantom under the guidance of a 1.5-T MR system using steady-state free precession (SSFP) sequencing. Various amounts of current were applied to the coils of the microcatheter to produce measureable tip deflections and navigate in vascular phantoms.
	The development of this device provides a platform for future testing and opportunity to revolutionize the endovascular interventional MRI environment.

Current applied to an endovascular microcatheter with microcoil tip made by laser lathe lithography can achieve controllable deflections under magnetic resonance (MR) guidance, which may improve speed and efficacy of navigation of vasculature during various endovascular procedures.

X-ray fluoroscopy-guided  endovascular procedures have several significant limitations, including  difficult catheter navigation and use of ionizing ...

Construction of a Preclinical Multimodality Phantom Using Tissue-mimicking Materials for Quality Assurance in Tumor Size Measurement

World Health Organization (WHO) and the Response Evaluation Criteria in Solid Tumors (RECIST) working groups advocated standardized criteria for radiologic assessment of solid tumors in response to anti-tumor drug therapy in the 1980s and 1990s, respectively. WHO criteria measure solid tumors in two-dimensions, whereas RECIST measurements use only one-dimension which is considered to be more reproducible 1, 2, 3,4,5. These criteria have been widely used as the only imaging biomarker approved by the United States Food and Drug Administration (FDA) 6. In order to measure tumor response to anti-tumor drugs on images with accuracy, therefore, a robust quality assurance (QA) procedures and corresponding QA phantom are needed.
To address this need, the authors constructed a preclinical multimodality (for ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI)) phantom using tissue-mimicking (TM) materials based on the limited number of target lesions required by RECIST by revising a Gammex US commercial phantom 7. The Appendix in Lee et al. demonstrates the procedures of phantom fabrication 7. In this article, all protocols are introduced in a step-by-step fashion beginning with procedures for preparing the silicone molds for casting tumor-simulating test objects in the phantom, followed by preparation of TM materials for multimodality imaging, and finally construction of the preclinical multimodality QA phantom. The primary purpose of this paper is to provide the protocols to allow anyone interested in independently constructing a phantom for their own projects. QA procedures for tumor size measurement, and RECIST, WHO and volume measurement results of test objects made at multiple institutions using this QA phantom are shown in detail in Lee et al. 8.

This paper describes in-house procedures of constructing a preclinical multimodality phantom made of tissue-mimicking (TM) materials for quality assurance (QA) of tumor size measurement in animal imaging modalities such as ultrasound (US), computed tomography (CT) and magnetic resonance imaging (MRI).

World Health Organization (WHO) and the Response Evaluation Criteria in Solid Tumors (RECIST) working groups advocated standardized criteria for ...

Hybrid &#181;CT-FMT imaging and image analysis

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to assess the biodistribution of novel probes and to assess disease progression using established molecular probes or reporter genes. The combination with an anatomical modality, e.g., micro computed tomography (&#181;CT), is beneficial for image analysis and for fluorescence reconstruction. We describe a protocol for multimodal &#181;CT-FMT imaging including the image processing steps necessary to extract quantitative measurements. After preparing the mice and performing the imaging, the multimodal data sets are registered. Subsequently, an improved fluorescence reconstruction is performed, which takes into account the shape of the mouse. For quantitative analysis, organ segmentations are generated based on the anatomical data using our interactive segmentation tool. Finally, the biodistribution curves are generated using a batch-processing feature. We show the applicability of the method by assessing the biodistribution of a well-known probe that binds to bones and joints.

Hybrid  &#181;CT-FMT imaging and image analysis

We describe a protocol for hybrid imaging, combining fluorescence-mediated tomography (FMT) with micro computed tomography (&#181;CT). After fusion and reconstruction, we perform interactive organ segmentation to extract quantitative measurements of the fluorescence distribution.

Fluorescence-mediated tomography (FMT) enables longitudinal and quantitative determination of the fluorescence distribution in vivo and can be used to ...

Custom-designed Laser-based Heating Apparatus for Triggered Release of Cisplatin from Thermosensitive Liposomes with Magnetic Resonance Image Guidance

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect resulting in significant reductions in systemic toxicity. Nonetheless, insufficient release of encapsulated drug from liposomes has limited their clinical efficacy. Temperature-sensitive liposomes have been engineered to provide site-specific release of drug in order to overcome the problem of limited tumor drug bioavailability. Our lab has designed and developed a heat-activated thermosensitive liposome formulation of cisplatin (CDDP), known as HTLC, to provide triggered release of CDDP at solid tumors. Heat-activated delivery in vivo was achieved in murine models using a custom-built laser-based heating apparatus that provides a conformal heating pattern at the tumor site as confirmed by MR thermometry (MRT). A fiber optic temperature monitoring device was used to measure the temperature in real-time during the entire heating period with online adjustment of heat delivery by alternating the laser power. Drug delivery was optimized under magnetic resonance (MR) image guidance by co-encapsulation of an MR contrast agent (i.e.,&#160;gadoteridol) along with CDDP into the thermosensitive liposomes as a means to validate the heating protocol and to assess tumor accumulation. The heating protocol consisted of a preheating period of 5 min prior to administration of HTLC and 20 min heating post-injection. This heating protocol resulted in effective release of the encapsulated agents with the highest MR signal change observed in the heated tumor in comparison to the unheated tumor and muscle. This study demonstrated the successful application of the laser-based heating apparatus for preclinical thermosensitive liposome development and the importance of MR-guided validation of the heating protocol for optimization of drug delivery.

AN MRI-compatible&#160;custom-designed laser-based heating apparatus has been developed to provide local heating of subcutaneous tumors in order to activate release of agents from thermosensitive liposomes specifically at the tumor region.

Liposomes have been employed as drug delivery systems to target solid tumors through exploitation of the enhanced permeability and retention (EPR) effect ...

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo

We demonstrate a superior method of 2D spectral-spatial imaging of stable radical reporter molecules at 250 MHz using rapid-scan electron-paramagnetic-resonance (RS-EPR), which can provide quantitative information under in vivo conditions on oxygen concentration, pH, redox status and concentration of signaling molecules (i.e., OH&#8226;, NO&#8226;). The RS-EPR technique has a higher sensitivity, improved spatial resolution (1 mm), and shorter acquisition time in comparison to the standard continuous wave (CW) technique. A variety of phantom configurations have been tested, with spatial resolution varying from 1 to 6 mm, and spectral width of the reporter molecules ranging from 16 &#181;T (160 mG) to 5 mT (50 G). A cross-loop bimodal resonator decouples excitation and detection, reducing the noise, while the rapid scan effect allows more power to be input to the spin system before saturation, increasing the EPR signal. This leads to a substantially higher signal-to-noise ratio than in conventional CW EPR experiments.

Rapid Scan Electron Paramagnetic Resonance Opens New Avenues for Imaging Physiologically Important Parameters In Vivo

A new electron paramagnetic resonance (EPR) method, rapid scan EPR (RS-EPR), is demonstrated for 2D spectral spatial imaging which is superior to the traditional continuous wave (CW) technique and opens new venues for in vivo imaging. Results are demonstrated at 250 MHz, but the technique is applicable at any frequency.

We demonstrate a superior method of 2D spectral-spatial imaging of stable radical reporter molecules at 250 MHz using rapid-scan ...

Metabolic Support of Excised, Living Brain Tissues During Magnetic Resonance Microscopy Acquisition

This protocol describes the procedures necessary to support normal metabolic functions of acute brain slice preparations during the collection of magnetic resonance (MR) microscopy data. While it is possible to perform MR collections on living, excised mammalian tissue, such experiments have traditionally been constrained by resolution limits and are thus incapable of visualizing tissue microstructure. Conversely, MR protocols that did achieve microscopic image resolution required the use of fixed samples to accommodate the need for static, unchanging conditions over lengthy scan times. The current protocol describes the first available MR technique that enables imaging of living, mammalian tissue samples at microscopic resolutions. Such data is of great importance to the understanding of how pathology-based contrast changes occurring at the microscopic level influence the content of macroscopic MR scans such as those used in the clinic. Once such an understanding is realized, diagnostic methods with greater sensitivity and accuracy can be developed, which will translate directly to earlier disease treatment, more accurate therapy monitoring and improved patient outcomes.
While the described methodology focuses on brain slice preparations, the protocol is adaptable to any excised tissue slice given that changes are made to the gas and perfusate preparations to accommodate the tissue&#39;s specific metabolic needs. Successful execution of the protocol should result in living, acute slice preparations that exhibit MR diffusion signal stability for periods up to 15.5 h. The primary advantages of the current system over other MR compatible perfusion apparatuses are its compatibility with the MR microscopy hardware required to attain higher resolution images and ability to provide constant, uninterrupted flow with carefully regulated perfusate conditions. Reduced sample throughput is a consideration with this design as only one tissue slice may be imaged at a time.

The current protocol describes a method by which users can maintain viability of acute hippocampal and cortical slice preparations during the collection of magnetic resonance microscopy data.

This protocol describes the procedures necessary to support normal metabolic functions of acute brain slice preparations during the collection of magnetic ...

Optimized Setup and Protocol for Magnetic Domain Imaging with In Situ Hysteresis Measurement

This paper elaborates the sample preparation protocols required to obtain optimal domain patterns using the Bitter method, focusing on the extra steps compared to standard metallographic sample preparation procedures. The paper proposes a novel bespoke rig for dynamic domain imaging with in situ BH (magnetic hysteresis) measurements and elaborates the protocols for the sensor preparation and the use of the rig to ensure accurate BH measurement. The protocols for static and ordinary dynamic domain imaging (without in situ BH measurements) are also presented. The reported method takes advantage of the convenience and high sensitivity of the traditional Bitter method and enables in situ BH measurement without interrupting or interfering with the domain wall movement processes. This facilitates establishing a direct and quantitative link between the domain wall movement processes&#8211;microstructural feature interactions in ferritic steels with their BH loops. This method is anticipated to become a useful tool for the fundamental study of microstructure&#8211;magnetic property relationships in steels and to help interpret the electromagnetic sensor signals for non-destructive evaluation of steel microstructures.

Optimized Setup and Protocol for Magnetic Domain Imaging with In Situ Hysteresis Measurement

This paper elaborates the sample and sensor preparation procedures and the protocols for using the test rig particularly for dynamic domain imaging with in situ BH measurements in order to achieve optimal domain pattern quality and accurate BH measurements.

This paper elaborates the sample preparation protocols required to obtain optimal domain patterns using the Bitter method, focusing on the extra steps ...

Measuring the Spin-Lattice Relaxation Magnetic Field Dependence of Hyperpolarized [1-13C]pyruvate

The fundamental limit to in vivo imaging applications of hyperpolarized 13C-enriched compounds is their finite spin-lattice relaxation times. Various factors affect the relaxation rates, such as buffer composition, solution pH, temperature, and magnetic field. In this last regard, the spin-lattice relaxation time can be measured at clinical field strengths, but at lower fields, where these compounds are dispensed from the polarizer and transported to the MRI, the relaxation is even faster and difficult to measure. To have a better understanding of the amount of magnetization lost during transport, we used fast field-cycling relaxometry, with magnetic resonance detection of 13C nuclei at ~0.75 T, to measure the nuclear magnetic resonance dispersion of the spin-lattice relaxation time of hyperpolarized [1-13C]pyruvate. Dissolution dynamic nuclear polarization was used to produce hyperpolarized samples of pyruvate at a concentration of 80 mmol/L and physiological pH (~7.8). These solutions were rapidly transferred to a fast field-cycling relaxometer so that relaxation of the sample magnetization could be measured as a function of time using a calibrated small flip angle (3&#176;-5&#176;). To map the T1 dispersion of the C-1 of pyruvate, we recorded data for different relaxation fields ranging between 0.237 mT and 0.705 T. With this information, we determined an empirical equation to estimate the spin-lattice relaxation of the hyperpolarized substrate within the mentioned range of magnetic fields. These results can be used to predict the amount of magnetization lost during transport and to improve experimental designs to minimize signal loss.

Measuring the Spin-Lattice Relaxation Magnetic Field Dependence of Hyperpolarized [1-13C]pyruvate

We present a protocol to measure the magnetic field dependence of the spin-lattice relaxation time of 13C-enriched compounds, hyperpolarized by means of dynamic nuclear polarization, using fast field-cycled relaxometry. Specifically, we have demonstrated this with [1-13C]pyruvate, but the protocol could be extended to other hyperpolarized substrates.

The fundamental limit to in vivo imaging applications of hyperpolarized 13C-enriched compounds is their finite spin-lattice relaxation times. Various ...