Name: in vivo epr assessment of ph, po2, redox status, and concentrations of phosphate and glutathione in the tumor microenvironment
Uploaded: 2018-03-16T13:00:00.000+00:00
Duration: 10 min 46 s
Description: Watch this Scientific Journal Video about In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment at JoVE.com

이 작품은 NIH에 의해 부분적으로 지원 된다 CA194013, CA192064 및 U54GM104942를 부여. WVCTSI VVK, AB, TDE를 시작에 대 한 인정 이다. 저자는 설명 실험 지원에 대 한 박사 M. Gencheva, K. 작성을 감사합니다. 내용은 전적으로 저자의 책임 이며 반드시 NIH의 공식 의견을 대표 하지 않는다.

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저자는 공개 없다.

제시 메서드는 중요 한 매개 변수 화학 TME의 즉 pO2, pH, 산화 환 원 상태 및 중간 Pi와 세포내 GSH의 농도의 비 침범 성 비보에 평가 대 한 수 있습니다. 낮은 분야 EPR, MRI와 자기 공명 기법, 비 침 투 적인 vivo에서 TME 매개 변수 프로 파일링에 대 한 선택의 방법이 있습니다. MRI는 해부학 적 구조를 시각화 하지만 기능 감도 부족. MRI, 달리 EPR 기술 기능 스핀 프로브와 함께 사?...

암 진행 및 치료에서 TME의 주요 역할은 점점 더 감사1. 단단한 종양, 조직 hypoxia2, 증3,4, 높은 감소 용량5, 세포내 GSH6,7, 높은 농도에서 TME의 중요 한 생리 적인 매개 변수 중 그리고 중간 Pi8 잘 문서화 되어있다. 비 침 투 적인 vivo에서 pO2, pH, Pi, GSH, 및 산화 환 원 평가 TME에서 생물 학적 과정에 대 한 독특한 통찰력을 제공 하 고 도와 항 암 약물과 치료 전략 TME 대상의 전 임상 심사에 대 한 사전 도구. 자기 공명 영상 (MRI) 및 낮은 필드 EPR 기반 기술에 의해 조직에 합리적인 알코올 관통은 그들이 이러한 TME 매개 변수의 비 침범 성 평가 대 한 가장 적절 한 접근 합니다. MRI 이미징 물 양성자에 크게 의존 하 고 해부학 해상도 제공 하기 위해 임상 설정에서 널리 이용 된다 하지만 기능 해상도 부족. Extracellular Pi 농도 pH 생 인산에서 신호에 따라 인-31 핵 자기 공명 (31P NMR) 측정 TME 특성화에 대 한 잠재적으로 매력 있지만 일반적으로 여러 번으로가 면 높은 세포내 Pi 농도9,10. 이, 달리 EPR 측정 분광학에 의존 하 고 특별히의 이미징 설계 상자성 프로브 기능 해상도 제공 하. 참고 exogenous EPR 프로브 외 인 한 이점을 NMR 프로브 EPR의 훨씬 더 높은 본질적인 감도와 내 생 적인 배경 EPR 신호의 부재 합니다. 듀얼 기능 pH 및 redox nitroxyl의 최근 개발11 프로브 및 다기능 trityl 프로브12 비보에 대 한 탁월한 기회 여러 TME 매개 변수의 동시 측정을 제공 하 고 그들의 상관 관계 분석 조사 배포 및 측정의 시간에 독립. 우리의 지식에 없는 다른 방법을 동시에 생체 내에서 생리 적으로 중요 한 화학 TME 매개 변수 pO2, pHe, Pi, redox, GSH 등 생활 주제에 평가를 사용할 수 있다.
에 대 한 조사 Vivo에서 기능 측정:
그림 1 상자성 프로브 TME 매개 변수, 미 립 자 및 녹는 프로브를 포함 하는 액세스 하는 데 사용의 화학 구조를 보여 줍니다. 고감도 기능, 조직 생활, 그리고 최소한의 독성 안정성 미 립 자 프로브 EPR 프로브가 비보에 대 한 수용 성 조사를 통해 선호 하는 몇 가지 이점이 있습니다. 예를 들어 입자 프로브는 몇 주 동안 조직 pO2 의 경도 측정에 대 한 허용 하는 녹는 프로브에 비해 조직 이식의 사이트에서 보존 배 증가 했습니다. 다른 한편으로, 녹는 프로브 EPR 기반 이미징 기법으로 수 있도록 여러 기능에서 부수적인 분석을 사용 하 여 해결 공간 측정을 제공 하 여 미 립 자 프로브를 능가할 (pO2, pH, Pi, 산화 환 원, 그리고 GSH)입니다.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56624/56624fig1.jpg"> 그림 1. TME 평가 분석 결과 조립 상자성 프로브의 화학 구조. 이 미 립 자 pO2 프로브, 링컨-BuO 포함 (R =-O (CH2)3채널3), 그리고 수용 성 조사: 듀얼 기능 pH 및 redox 프로브, NR; GSH-민감한 프로브, RSSR; 다기능 pO2와 pH, 그리고 extracellular microenvironment, 희망 조사. 의 Pi 프로브 이러한 프로브 합성 제공된 참조 11,12에서 설명 하고있다. <a href="//cloudflare.jove.com/files/ftp_upload/56624/56624fig1large.jpg" target="_blank">이 그림의 더 큰 버전을 보려면 여기를 클릭 하십시오.</a>

<table><tbody><tr><td>L-band EPR spectrometer</td><td>Magnettech, Germany</td><td></td><td>L-band (1.2 GHz) electron paramagnetic resonance (EPR) spectrometer for collection in vitro and in vivo spectra of paramagnetic molecules</td></tr><tr><td>&amp;nbsp;Temperature &amp;amp; Gas Controller&amp;nbsp;</td><td>Noxygen, Germany</td><td></td><td>Temperature &amp;amp; Gas Controller designed to control and adjust the temperature and gas composition&amp;nbsp;&amp;nbsp;</td></tr><tr><td>Sonicator</td><td>Fisher Scientific</td><td></td><td></td></tr><tr><td>GSH (L-Glutathione reduced)</td><td>Sigma-Aldrich</td><td>G4251</td><td></td></tr><tr><td>MMTV-PyMT&amp;nbsp; mice</td><td>In house</td><td></td><td></td></tr><tr><td>DMEM</td><td>Thermo Fisher Scientific</td><td>11995065</td><td></td></tr><tr><td>Met-1 murine breast cancer cells</td><td>In house</td><td></td><td></td></tr><tr><td>C57Bl/6 wild type mice&amp;nbsp;</td><td>Jackson Laboratory</td><td></td><td></td></tr><tr><td>Trypsin</td><td>Thermo Fisher Scientific</td><td>25200056</td><td></td></tr><tr><td>Trypan Blue Exclusion Dye&amp;nbsp;</td><td>Thermo Fisher Scientific</td><td>T10282</td><td></td></tr><tr><td>Ohmeda Fluotec 3&amp;nbsp;</td><td></td><td></td><td></td></tr><tr><td>Isoflurane (IsoFlo)</td><td>Abbott Laboratories</td><td></td><td></td></tr><tr><td>Sodium phosphate dibasic</td><td>Sigma-Aldrich</td><td>S9763</td><td></td></tr><tr><td>Sodium phosphate monobasic</td><td>sigma-Aldrich</td><td>S07051</td><td></td></tr><tr><td>Sodium Chloride</td><td>sigma-Aldrich</td><td>S7653</td><td></td></tr><tr><td>Hydrochloric acid</td><td>sigma-Aldrich</td><td>320331</td><td></td></tr><tr><td>Sodium Hydroxide</td><td>sigma-Aldrich</td><td>S8045</td><td></td></tr><tr><td>Glucose</td><td>sigma-Aldrich</td><td></td><td></td></tr><tr><td>Glucose oxydase</td><td>sigma-Aldrich</td><td></td><td></td></tr><tr><td>Lauda Circulator E100</td><td>Lauda-Brikmann</td><td></td><td></td></tr><tr><td>pH meter Orion</td><td>Thermo Scientific&amp;nbsp;</td><td></td><td></td></tr><tr><td>LiNc-BuO probe</td><td>In house</td><td></td><td>The Octa-n-Butoxy-Naphthalocyanine probe was synthesizided according to ref 13</td></tr><tr><td>NR probe</td><td>In house</td><td></td><td>The Nitroxide probe was synthesizided according to ref 11</td></tr><tr><td>RSSR probe</td><td>In house</td><td></td><td>The di-Nitroxide probe was synthesizided according to ref 15</td></tr><tr><td>HOPE &lt;em&gt;probe&lt;/em&gt;</td><td>In house</td><td></td><td>The monophoshonated Triarylmethyl probe was synthesizided according to ref 12</td></tr></tbody></table>

in vivo epr assessment of ph, po2, redox status, and concentrations of phosphate and glutathione in the tumor microenvironment

이 프로토콜 낮은 필드 전자 상자성 공명 (EPR)의 기능을 보여줍니다-화학 종양 microenvironment (TME)에 정량적 인 정보를 제공 하는 기능 상자성 프로브 조합에서 기법을 기반으로 포함 p O2, pH, 산화 환 원 상태, 간 질 성 무기 인산 (Pi), 그리고 세포내 티 (GSH)의 농도. 특히, 최근에 개발 된 수용 성 다기능 trityl 프로브 응용 프로그램 탁월한 기회를 제공 한다 vivo에서 pH, pO2 와 P의 동시 측정 e에서 나 xtracellular 공간 (희망 조사)입니다. 세 개의 매개 변수는 단일 프로브를 사용 하 여 측정 프로브 배포 및 시간 측정의 그들의 상관 분석에 대 한 수 있습니다.

조직 p O 2 평가 링컨-BuO를 사용 하 여 조사:
우리는 갓된 링컨 BuO microcrystals 서 스 펜 션의 교정 수행 단계 1.1 아래 설명 된 절차를 사용 하 여. 그림 2 는 일반적인 산소 의존 링컨 BuO 프로브 여성 C57Bl/6 마우스의 세포를 주입 하 ...

Watch this Scientific Journal Video about In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment at JoVE.com

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

낮은 분야 (L-밴드, 1.2 g h z) 전자 상자성 공명 녹는 nitroxyl 및 trityl 프로브를 사용 하 여 유방암의 마우스 모델에서 종양 microenvironment에 생리 적으로 중요 한 매개 변수 평가 대 한 시연입니다.

Vivo에서 PH, pO2, 산화 환 원 상태, 고 농도의 인산 염 및 종양 Microenvironment에 티의 EPR 평가

This protocol demonstrates the capability of low-field electron paramagnetic resonance (EPR)-based techniques in combination with functional paramagnetic ...

in-vivo-epr-assessment-ph-po2-redox-status-concentrations-phosphate

Research

JoVE Journal

Cancer Research

In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

8.2K 조회수.  West Virginia University. 낮은 분야 (L-밴드, 1.2 g h z) 전자 상자성 공명 녹는 nitroxyl 및 trityl 프로브를 사용 하 여 유방암의 마우스 모델에서 종양 microenvironment에 생리 적으로 중요 한 매개 변수 평가 대 한 시연입니다.

비디오: In Vivo EPR Assessment of pH, pO2, Redox Status, and Concentrations of Phosphate and Glutathione in the Tumor Microenvironment

Hyperpolarized 13C Metabolic Magnetic Resonance Spectroscopy and Imaging

In the past decades, new methods for tumor staging, restaging, treatment response monitoring, and recurrence detection of a variety of cancers have emerged in conjunction with the state-of-the-art positron emission tomography with 18F-fluorodeoxyglucose ([18F]-FDG PET). 13C magnetic resonance spectroscopic imaging (13CMRSI) is a minimally invasive imaging method that enables the monitoring of metabolism in vivo and in real time. As with any other method based on 13C nuclear magnetic resonance (NMR), it faces the challenge of low thermal polarization and a subsequent low signal-to-noise ratio due to the relatively low gyromagnetic ratio of 13C and its low natural abundance in biological samples. By overcoming these limitations, dynamic nuclear polarization (DNP) with subsequent sample dissolution has recently enabled commonly used NMR and magnetic resonance imaging (MRI) systems to measure, study, and image key metabolic pathways in various biological systems. A particularly interesting and promising molecule used in 13CMRSI is [1-13C]pyruvate, which, in the last ten years, has been widely used for in vitro, preclinical, and, more recently, clinical studies to investigate the cellular energy metabolism in cancer and other diseases. In this article, we outline the technique of dissolution DNP using a 3.35 T preclinical DNP hyperpolarizer and demonstrate its usage in in vitro studies. A similar protocol for hyperpolarization may be applied for the most part in in vivo studies as well. To do so, we used lactate dehydrogenase (LDH) and catalyzed the metabolic reaction of [1-13C]pyruvate to [1-13C]lactate in a prostate carcinoma cell line, PC3, in vitro using 13CMRSI.

Hyperpolarized 13C Metabolic Magnetic Resonance Spectroscopy and Imaging

Dynamic nuclear polarization with subsequent sample dissolution has enabled real-time studies of metabolism in biological systems. Hyperpolarized [1-13C]pyruvate was used to study lactate dehydrogenase activity in a prostate carcinoma cell line in vitro.

과분극<sup&gt; (13)</sup&gt; C 대사 자기 공명 분광학 및 이미징

In the past decades, new methods for tumor staging, restaging, treatment response monitoring, and recurrence detection of a variety of cancers have ...

연구

암 연구학

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

의학

Use of Electron Paramagnetic Resonance in Biological Samples at Ambient Temperature and 77 K

The accurate and specific detection of reactive oxygen species (ROS) in different cellular and tissue compartments is essential to the study of redox-regulated signaling in biological settings. Electron paramagnetic resonance spectroscopy (EPR) is the only direct method to assess free radicals unambiguously. Its advantage is that it detects physiologic levels of specific species with a high specificity, but it does require specialized technology, careful sample preparation, and appropriate controls to ensure accurate interpretation of the data. Cyclic hydroxylamine spin probes react selectively with superoxide or other radicals to generate a nitroxide signal that can be quantified by EPR spectroscopy. Cell-permeable spin probes and spin probes designed to accumulate rapidly in the mitochondria allow for the determination of superoxide concentration in different cellular compartments.
In cultured cells, the use of cell permeable 1-hydroxy-3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine (CMH) along with and without cell-impermeable superoxide dismutase (SOD) pretreatment, or use of cell-permeable PEG-SOD,&#160;allows for the differentiation of extracellular from cytosolic superoxide. The mitochondrial 1-hydroxy-4-[2-triphenylphosphonio)-acetamido]-2,2,6,6-tetramethyl-piperidine,1-hydroxy-2,2,6,6-tetramethyl-4-[2-(triphenylphosphonio)acetamido] piperidinium dichloride (mito-TEMPO-H) allows for measurement of mitochondrial ROS (predominantly superoxide).
Spin probes and EPR spectroscopy can also be applied to in vivo models. Superoxide can be detected in extracellular fluids such as blood and alveolar fluid, as well as tissues such as lung tissue. Several methods are presented to process and store tissue for EPR measurements and deliver intravenous 1-hydroxy-3-carboxy-2,2,5,5-tetramethylpyrrolidine (CPH) spin probe in vivo. While measurements can be performed at room temperature, samples obtained from in vitro and in vivo models can also be stored at -80 &#176;C and analyzed by EPR at 77 K. The samples can be stored in specialized tubing stable at -80 &#176;C and run at 77 K to enable a practical, efficient, and reproducible method that facilitates storing and transferring samples.

Electron paramagnetic resonance (EPR) spectroscopy is an unambiguous method to measure free radicals. The use of selective spin probes allows for detection of free radicals in different cellular compartments. We present a practical, efficient method to collect biological samples that facilitate treating, storing, and transferring samples for EPR measurements.

The accurate and specific detection of reactive oxygen species (ROS) in different cellular and tissue compartments is essential to the study of ...

생화학

Isolation of Proximal Fluids to Investigate the Tumor Microenvironment of Pancreatic Adenocarcinoma

Pancreatic adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death, and soon to become the second. There is an urgent need of variables associated to specific pancreatic pathologies to help preoperative differential diagnosis and patient profiling. Pancreatic juice is a relatively unexplored body fluid, which, due to its close proximity to the tumor site, reflects changes in the surrounding tissue. Here we describe in detail the intraoperative collection procedure. Unfortunately, translating pancreatic juice collection to murine models of PDAC, to perform mechanistic studies, is technically very challenging. Tumor interstitial fluid (TIF) is the extracellular fluid, outside blood and plasma, which bathes tumor and stromal cells. Similarly to pancreatic juice, for its property to collect and concentrate molecules that are found diluted in plasma, TIF can be exploited as an indicator of microenvironmental alterations and as a valuable source of disease-associated biomarkers. Since TIF is not readily accessible, various techniques have been proposed for its isolation. We describe here two simple and technically undemanding methods for its isolation: tissue centrifugation and tissue elution.

Pancreatic juice is a precious source of biomarkers for human pancreatic cancer. We describe here a method for intraoperative collection procedure. To overcome the challenge of adopting this procedure in murine models, we suggest an alternative sample, tumor interstitial fluid, and describe here two protocols for its isolation.

췌장 선암의 종양 미세 환경을 조사하기위한 근위액 분리

Pancreatic adenocarcinoma (PDAC) is the fourth leading cause of cancer-related death, and soon to become the second. There is an urgent need of variables ...

Probing Metabolism and Viscosity of Cancer Cells using Fluorescence Lifetime Imaging Microscopy

Viscosity is an important physical property of a biological membrane, as it is one of the key parameters for the regulation of morphological and physiological state of living cells. Plasma membranes of tumor cells are known to have significant alterations in their composition, structure, and functional characteristics. Along with dysregulated metabolism of glucose and lipids, these specific membrane properties help tumor cells to adapt to the hostile microenvironment and develop resistance to drug therapies. Here, we demonstrate the use of fluorescence lifetime imaging microscopy (FLIM) to sequentially image cellular metabolism and plasma membrane viscosity in live cancer cell culture. Metabolic assessments are performed by detecting fluorescence of endogenous metabolic cofactors, such as reduced nicotinamide adenine dinucleotide NAD(P)H and oxidized flavins. Viscosity is measured using a fluorescent molecular rotor, a synthetic viscosity-sensitive dye, with a strong fluorescence lifetime dependence on the viscosity of the immediate environment. In combination, these techniques enable us to better understand the links between membrane state and metabolic profile of cancer cells and to visualize the changes induced by chemotherapy.

Here, we demonstrate the use of fluorescence lifetime imaging microscopy (FLIM) to sequentially image cellular metabolism and plasma membrane viscosity in live cancer cell culture. Metabolic assessments are performed by detecting endogenous fluorescence. Viscosity is measured using a fluorescent molecular rotor.

Viscosity is an important physical property of a biological membrane, as it is one of the key parameters for the regulation of morphological and ...

Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens

Pseudomyxoma peritonei (PMP) is a rare condition that results from the dissemination of a mucinous primary tumor and the resultant accumulation of mucin-secreting tumor cells in the peritoneal cavity. PMP can arise from various types of cancers, including appendiceal, ovarian, and colorectal, though appendiceal neoplasms are by far the most common etiology. PMP is challenging to study due to its (1) rarity, (2) limited murine models, and (3) mucinous, acellular histology. The method presented here allows real-time visualization and interrogation of these tumor types using patient-derived ex vivo organotypic slices in a preparation where the tumor microenvironment (TME) remains intact. In this protocol, we first describe the preparation of tumor slices using a vibratome and subsequent long-term culture. Second, we describe confocal imaging of tumor slices and how to monitor functional readouts of viability, calcium imaging, and local proliferation. In short, slices are loaded with imaging dyes and are placed in an imaging chamber that can be mounted onto a confocal microscope. Time-lapse videos and confocal images are used to assess the initial viability and cellular functionality. This procedure also explores translational cellular movement, and paracrine signaling interactions in the TME. Lastly, we describe a dissociation protocol for tumor slices to be used for flow cytometry analysis. Quantitative flow cytometry analysis can be used for bench-to-bedside therapeutic testing to determine changes occurring within the immune landscape and epithelial cell content.

Culture and Imaging of Ex Vivo Organotypic Pseudomyxoma Peritonei Tumor Slices from Resected Human Tumor Specimens

We describe a protocol for the production, culture, and visualization of human cancers, which have metastasized to the peritoneal surfaces. Resected tumor specimens are cut using a vibratome and cultured on permeable inserts for increased oxygenation and viability, followed by imaging and downstream analyses using confocal microscopy and flow cytometry.

절제된 인간 종양 표본에서 생체 외 유기형 가성 점액종 복막 종양 절편의 배양 및 이미징

Pseudomyxoma peritonei (PMP) is a rare condition that results from the dissemination of a mucinous primary tumor and the resultant accumulation of ...

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.

빠른 스캔 전자 스핀 공명 이미징 생리 학적으로 중요 매개 변수에 대한 새로운 길을 엽니 다<em&gt; 생체</em

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

생체공학

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both direct and rapid analysis of therapeutic sensitivity and resistance. However, recent evidence suggests that therapeutic response is not exclusive to the inherent molecular composition of cancer cells but rather is greatly influenced by the tumor cell microenvironment, a feature that cannot be recapitulated by traditional culturing methods. Even implementation of tumor xenografts, though providing a wealth of information on drug delivery/efficacy, cannot capture the tumor cell/microenvironment crosstalk (i.e., soluble factors) that occurs within human tumors and greatly impacts tumor response. To this extent, we have developed an ex vivo (fresh tissue sectioning) technique which allows for the direct assessment of treatment response for preclinical and clinical therapeutics development. This technique maintains tissue integrity and cellular architecture within the tumor cell/microenvironment context throughout treatment response providing a more precise means to assess drug efficacy.

Ex Vivo Treatment Response of Primary Tumors and/or Associated Metastases for Preclinical and Clinical Development of Therapeutics

Established cancer cell lines and xenografts have been the mainstay of cancer research for the past several decades. However, recent evidence suggests that therapeutic response is greatly influenced by the tumor cell microenvironment. Therefore, we have developed an ex vivo analysis of primary tumor specimens for drug development purposes.

기본 종양 및 / 또는 전임상에 대한 관련 전이 및 치료제의 임상 개발의 전 생체 치료 응답

The molecular analysis of established cancer cell lines has been the mainstay of cancer research for the past several decades. Cell culture provides both ...

Guided Tumor Inoculation and Multimodal Imaging in Mice

A Dorsal Skinfold Window Chamber Tumor Mouse Model for Combined Intravital Microscopy and Magnetic Resonance Imaging in Translational Cancer Research

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the tumor microenvironment and its response to therapy. These imaging modalities have micron-scale resolution but have limited use in the clinic due to their shallow penetration depth into tissue. More clinically applicable imaging modalities such as CT, MRI, and PET have much greater penetration depth but have comparatively lower spatial resolution (mm scale). 
To translate preclinical intravital imaging findings into the clinic, new methods must be developed to bridge this micro-to-macro resolution gap. Here we describe a dorsal skinfold window chamber tumor mouse model designed to enable preclinical intravital and clinically applicable (CT and MR) imaging in the same animal, and the image analysis platform that links these two disparate visualization methods. Importantly, the described window chamber approach enables the different imaging modalities to be co-registered in 3D using fiducial markers on the window chamber for direct spatial concordance. This model can be used for validation of existing clinical imaging methods, as well as for the development of new ones through direct correlation with "ground truth" high-resolution intravital findings. 
Finally, the tumor response to various treatments-chemotherapy, radiotherapy, photodynamic therapy-can be monitored longitudinally with this methodology using preclinical and clinically applicable imaging modalities. The dorsal skinfold window chamber tumor mouse model and imaging platforms described here can thus be used in a variety of cancer research studies, for example, in translating preclinical intravital microscopy findings to more clinically applicable imaging modalities such as CT or MRI.

저자 스포트라이트: 고해상도 생체 내 영상 및 MRI를 통합하여 정위 신체 방사선 치료 계획 향상

Translation of Intravital microscopy findings is challenged by its shallow depth penetration into tissue. Here we describe a dorsal window chamber mouse model that enables co-registration of intravital microscopy and clinically applicable imaging modalities (e.g., CT, MRI) for direct spatial correlation, potentially streamlining clinical translation of intravital microscopy findings.

중개 암 연구에서 생체 내 현미경 및 자기 공명 영상의 결합을 위한 Dorsal Skinfold Window Chamber 종양 마우스 모델

Preclinical intravital imaging such as microscopy and optical coherence tomography have proven to be valuable tools in cancer research for visualizing the ...

Vivo에서 PH, pO₂, 산화 환 원 상태, 고 농도의 인산 염 및 종양 Microenvironment에 티의 EPR 평가

요약

더 많은 비디오 탐색

이 비디오의 챕터

과분극

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

Use of Electron Paramagnetic Resonance in Biological Samples at Ambient Temperature and 77 K

췌장 선암의 종양 미세 환경을 조사하기위한 근위액 분리

Probing Metabolism and Viscosity of Cancer Cells using Fluorescence Lifetime Imaging Microscopy

절제된 인간 종양 표본에서 생체 외 유기형 가성 점액종 복막 종양 절편의 배양 및 이미징

빠른 스캔 전자 스핀 공명 이미징 생리 학적으로 중요 매개 변수에 대한 새로운 길을 엽니 다

기본 종양 및 / 또는 전임상에 대한 관련 전이 및 치료제의 임상 개발의 전 생체 치료 응답

중개 암 연구에서 생체 내 현미경 및 자기 공명 영상의 결합을 위한 Dorsal Skinfold Window Chamber 종양 마우스 모델

중개 암 연구에서 생체 내 현미경 및 자기 공명 영상의 결합을 위한 Dorsal Skinfold Window Chamber 종양 마우스 모델