eoe sub articles

EoE Sub Articles

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. In Vivo PET Imaging with 89Zr-DFO-J591
CAUTION: This step of the protocol involves the handling and manipulation of radioactivity. Before performing these steps, researchers should consult with their home institution&#39;s Radiation Safety Department. All possible steps should be taken to minimize exposure to ionizing radiation.
<ol>
	<li>In male athymic nude mice, subcutaneously implant 5 x 106 lymph node carcinoma of the prostate (LNCaP) cancer cells and allow these to grow to a 100-150 mm3 xenograft (3-4 weeks after inoculation).</li>
	<li>Dilute the 89Zr-DFO-J591 radioimmunoconjugate to a concentration of 1.0 mCi/ml in 0.9% sterile saline.</li>
	<li>Inject 200 &#181;l of the 89Zr-DFO-J591 (DFO = desferrioxamine, J591 =&#160; prostate-specific membrane antigen [PSMA] targeting antibody) solution (200 &#181;Ci; 7.4 MBq) into the lateral tail vein of the xenograft-bearing mice.</li>
	<li>At the desired imaging time point (e.g., 12, 24, 48, 72, 96, or 120 hr post-injection), anesthetize the mouse with a 2% isoflurane: oxygen gas mixture.</li>
	<li>Place the mouse on the bed of the small animal PET scanner and maintain anesthesia during the scan using a 1% isoflurane: oxygen gas mixture. Prior to placing the animal on the scanner bed, verify anesthesia using the toe-pinch method and apply ophthalmic ointment to the eyes of the mouse to prevent drying during anesthesia.</li>
	<li>Acquire the PET data for the mouse via a static scan with a minimum of 40 million coincident events using an energy window of 350-700 keV and a coincidence timing window of 6 nsec.</li>
	<li>After completing the acquisition of the image, do not leave the mouse unattended, and do not place it in a cage with other mice until it has regained consciousness.</li>
</ol>

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>p-SCN-Bn-DFO</td>
			<td>Macrocyclics</td>
			<td>B-705</td>
			<td>Store at -80 &#176;C</td>
		</tr>
		<tr>
			<td>[89Zr]Zr-oxalate</td>
			<td>Various, including Perkin-Elmer</td>
			<td>-</td>
			<td>Caution: Radioactive material</td>
		</tr>
	</tbody>
</table>

an in vivo pet imaging technique to detect tumors in a murine model using radiolabeled antibodies

Source: Zeglis, B. M., et al. <a href="https://app-jove-com.remotexs.ntu.edu.sg/v/52521">The Bioconjugation and Radiosynthesis of 89Zr-DFO-labeled Antibodies.</a> J. Vis. Exp. (2015).
This video demonstrates a technique involving a radioimmunoconjugate for the in vivo imaging of a tumor in a mouse model. Upon injection of the radioimmunoconjugate, the antibodies of the conjugate bind to a specific membrane antigen on the cancer cells. The radiotracer of the conjugate emits positrons that interact with neighboring electrons to produce energy, which is detected by a positron emission tomography (PET) scanner to create an image of the tumor.

An In Vivo PET Imaging Technique to Detect Tumors in a Murine Model Using Radiolabeled Antibodies

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. In ...

Take prostate cancer cells and inject them subcutaneously into an anesthetized, &#160;immunocompromised nude mouse.
The lack of an immune response allows the injected cells to survive and multiply, forming a tumor.
The tumor cells overexpress a prostate-specific membrane antigen or PSMA, a cell-surface protein.
Take PSMA-specific antibodies conjugated to a radiotracer and inject them intravenously into the tail of the tumor-bearing mouse.
Antibodies reach the tumor cells and bind to PSMA, labeling the tumor.
Place the anesthetized mouse in a positron-emission tomography or PET scanner and obtain a scan.
The produced image shows the distribution of the radiotracer, highlighting areas of high antibody uptake, which correspond to the tumor location.

an-vivo-pet-imaging-technique-to-detect-tumors-murine-model-using

Nanyang Technological University

Research

JoVE Encyclopedia of Experiments

Immunology

Antibody-Based Technologies

Antibody-Based Imaging and Detection Methods

94 Views. Source: Zeglis, B. M., et al. The Bioconjugation and Radiosynthesis of 89Zr-DFO-labeled Antibodies. J. Vis. Exp. (2015). This video demonstrates a technique involving a radioimmunoconjugate for the in vivo imaging of a tumor in a mouse model. Upon injection of the radioimmunoconjugate, the antibodies of the conjugate bind to a specific membrane antigen on the cancer cells. The radiotracer of the conjugate emits positrons that interact with neighboring electrons to produce energy, which is detec...

Video: An In Vivo PET Imaging Technique to Detect Tumors in a Murine Model Using Radiolabeled Antibodies - Concept

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models

Metastasis is responsible for most cancer deaths. Despite extensive research, the mechanistic understanding of the complex processes governing metastasis remains incomplete. In vivo models are paramount for metastasis research, but require refinement. Tracking spontaneous metastasis by non-invasive in vivo imaging is now possible, but remains challenging as it requires long-time observation and high sensitivity. We describe a longitudinal combined radionuclide and fluorescence whole-body in vivo imaging approach for tracking tumor progression and spontaneous metastasis. This reporter gene methodology employs the sodium iodide symporter (NIS) fused to a fluorescent protein (FP). Cancer cells are engineered to stably express NIS-FP followed by selection based on fluorescence-activated cell sorting. Corresponding tumor models are established in mice. NIS-FP expressing cancer cells are tracked non-invasively in vivo at the whole-body level by positron emission tomography (PET) using the NIS radiotracer [18F]BF4-. PET is currently the most sensitive in vivo imaging technology available at this scale and enables reliable and absolute quantification. Current methods either rely on large cohorts of animals that are euthanized for metastasis assessment at varying time points, or rely on barely quantifiable 2D imaging. The advantages of the described method are: (i) highly sensitive non-invasive in vivo 3D PET imaging and quantification, (ii) automated PET tracer production, (iii) a significant reduction in required animal numbers due to repeat imaging options, (iv) the acquisition of paired data from subsequent imaging sessions providing better statistical data, and (v) the intrinsic option for ex vivo confirmation of cancer cells in tissues by fluorescence microscopy or cytometry. In this protocol, we describe all steps required for routine NIS-FP-afforded non-invasive in vivo cancer cell tracking using PET/CT and ex vivo confirmation of in vivo results. This protocol has applications beyond cancer research whenever in vivo localization, expansion and long-time monitoring of a cell population is of interest.

We describe a protocol for preclinical in vivo tracking of cancer metastasis. It is based on a radionuclide-fluorescence reporter combining the sodium iodide symporter, detected by non-invasive [18F]tetrafluoroborate-PET, and a fluorescent protein for streamlined ex vivo confirmation. The method is applicable for preclinical in vivo cell tracking beyond tumor biology.

Metastasis is responsible for most cancer deaths. Despite extensive research, the mechanistic understanding of the complex processes governing metastasis ...

JoVE Journal

Cancer Research

A Practical Guide for the Production and PET/CT Imaging of 68Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Neuroendocrine tumors are a rare form of cancer that arise from neuroendocrine cells and can be present at almost any location throughout the body. Although heterogeneous in presentation, a common denominator among these tumors is the overexpression of somatostatin receptors. 68Ga-DOTATATE is a somatostatin analog labeled with the positron emitter gallium-68 (68Ga). For well-differentiated neuroendocrine tumors, 68Ga-DOTATATE positron emission tomography (PET)/computed tomography (CT) imaging is used for diagnosis, determination of disease burden, and therapy selection.
This protocol details the radiolabeling of 68Ga-DOTATATE, quality control, patient preparation, and subsequent PET/CT imaging. Radiolabeling of 68Ga-DOTATATE is performed with a fully automated labeling module coupled to a germanium-68 (68Ge)/68Ga generator. Quality control of the final product evaluates radiochemical purity with instant thin-layer chromatography and solid-phase chromatography, and pH prior to patient injection. Periodic quality control is performed to determine 68Ge breakthrough, sterility, and (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) content. Patient preparation includes patient instructions, a protocol for 68Ga-DOTATATE during treatment with somatostatin analogs, and intravenous administration of the radiopharmaceutical. For PET/CT imaging, the acquisition and reconstruction settings are described. For each step, radiation safety will be highlighted, as well as time constrictions due to the short half-life of 68Ga.
Fully automated in-house production and quality control of 68Ga-DOTATATE leads to very high success rates (95%) and produces two to four patient dosages per batch, depending on the yield of the generator. In conclusion, 68Ga-DOTATATE PET/CT imaging is a noninvasive and fast method of providing information on the tumor burden of neuroendocrine tumors (NETs) while also assisting in diagnosis and therapy selection.

A Practical Guide for the Production and PET/CT Imaging of 68Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Well-differentiated neuroendocrine tumors overexpress somatostatin receptors which can be utilized for diagnostic imaging with the radiolabeled somatostatin analog 68Ga-DOTATATE. This protocol details the radiolabeling of 68Ga-DOTATATE, quality control, patient preparation, and subsequent PET/CT imaging. Radiation safety and time constrictions due to the short half-life of 68Ga are taken into account.

Neuroendocrine tumors are a rare form of cancer that arise from neuroendocrine cells and can be present at almost any location throughout the body. ...

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and evaluate novel therapeutic approaches. Non-invasive whole-body imaging using positron emission tomography (PET) provides critical insights into the in vivo characteristics of tissues at the molecular level in real-time. We present here a protocol for orthotopic HCC xenograft creation with and without hepatic artery ligation (HAL) to induce tumor hypoxia and the assessment of their tumor metabolism in vivo using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG) PET/magnetic resonance (MR) imaging. Tumor hypoxia could be readily visualized using the hypoxia marker [18F]FMISO, and it was found that the [18F]FMISO uptake was higher in HCC mice that underwent HAL than in the non-HAL group, whereas [18F]FDG could not distinguish tumor hypoxia between the two groups. HAL tumors also displayed a higher level of hypoxia-inducible factor (HIF)-1&#945; expression in response to hypoxia. Quantification of HAL tumors showed a 2.3-fold increase in [18F]FMISO uptake based on the standardized value uptake (SUV) approach.

Here, we present a protocol to create orthotopic hepatocellular carcinoma xenografts with and without hepatic artery ligation and perform non-invasive positron emission tomography (PET) imaging of tumor hypoxia using [18F]Fluoromisonidazole ([18F]FMISO) and [18F]Fluorodeoxyglucose ([18F]FDG).

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

An In Vivo PET Imaging Technique to Detect Tumors in a Murine Model Using Radiolabeled Antibodies

Transcript

An In Vivo PET Imaging Technique to Detect Tumors in a Murine Model Using Radiolabeled Antibodies

Radionuclide-fluorescence Reporter Gene Imaging to Track Tumor Progression in Rodent Tumor Models

A Practical Guide for the Production and PET/CT Imaging of ⁶⁸Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma