Boyuna görüntüleme bir hastalığın ilerleyişini takip ya da tedavi edici bir etkisi ölçmek için klinik öncesi çalışmalarda kullanılır. Onkoloji, optik yöntemler, tümör büyümesini izlemek ve böyle bir çalışmanın her zaman noktasında, hücre büyümesi veya gen ifadesi kesin kantitatif sunmak için titiz araçları sağlar. MicroCT gibi yüksek çözünürlüklü bir tekniği kullanılarak ölçülür Anatomik değişiklikler olabilir, ama Boyuna görüntüleme için düşük bir X-ışını doz biyolojik bozulmaları önlemek için kullanılması gerekir. Optik ve microCT görüntüler maksimum bilgi hayvan modeli elde edilir emin fonksiyonel ve anatomik veri bir arada sağlamak için ortak tescil edilebilir. 1. Hücre hazırlık <ol><li> Kaliper fare modelleri, ön-klinik araştırma lusiferaz ifade kanser hücre hatları bir dizi sağlar. </li><li> MDA-MB-231-luc-D2H2LN, in vivo tumorgenesis optik bir göstergesi olarak kullanılabilir lusiferaz gen ifade eden bir insan meme tümör hücre hattı. Bu hücre hattı D3H1 meme yağ yastığı tümör kaynaklanan kendiliğinden bir lenf nodu metastazı oluşturulur ve agresif metastaz formu bilinmektedir. </li><li> Hücreleri kolayca seçim işaretleyicilerini gerek standart medya yetişen bir patojen içermeyen dondurulmuş kültür olarak verilmektedir. </li><li> Enjeksiyondan önce hayvan Lusiferaz aktivitesi doğrulamak için% 90 konfluent balon trypsinization hasat edilir. Lusiferaz aktivitesi mikrotiter plate 50.000 hücreleri dağıtım ve seri seyreltme performans ile ölçülür. </li></ol> 2. Hayvanlarda hücrelerin intrakardiyak enjeksiyon <ol><li> Enjeksiyondan önce, hayvanlar% 3 izofluran kullanarak anestezi vardır. </li><li> Sol ventrikül 50ul hacmi 1-3 milyon hücre enjekte edilir. Hücre süspansiyon 150ug/ml D-luciferase enjeksiyon tekniği doğrulamak için içerir. </li><li> IVIS sistemi doğru bir intrakardiyak enjeksiyon fareler onaylamak için görüntülü. Vücutta BLI sinyal göstermiyor Fare doğru enjekte edilmiştir. Sinyal lokalize ise kalp hücrelerinin doğru enjekte değildi. </li></ol> 3. BLI görüntüleme metastaz izlemek için <ol><li> Görüntüleme servis IVIS Spektrum ve QuantumFX microCT sistemi hem de veri yakalamak için fare destekler. </li><li> Mekik hayvan görüntüleme için aynı pozisyonda tutmanıza yardımcı olur ve co-kayıt otomatik yardım fiducial sinyaller içermektedir. Odasına anestezi korumak için isofluorance buharlaştırıcı bağlı. </li><li> Yaşayan Image yazılımı 4.1 Görüntüleme sihirbazı otomatik pozlama süresi, f-stop ve binning seçin ve nicel veri toplama sağlamak için kullanılabilir. </li><li> Çoklu görüntüleri elde ve deney doğa bağlı olarak saniye veya ay kapsayan boylamsal çalışmalar mukayese edilebilir. </li><li> Gün 6 post enjeksiyon hücre, BLI görüntüleme IVIS Spectrum dokuz fareler yedi kemik metastazı gösterir. </li></ol> 4. Mikro BT kullanarak osteolitik lezyonlar Boyuna izlenmesi <ol><li> Kuantum FX mikro CT sistemi BLI kullanılarak görülebilir net kemik metastazları gösterdi Fare görüntülenmiştir. </li><li> Kuantum FX mikro BT sistemi preklinik çalışmalarda gerçek boyuna mikro BT yetecek kadar düşük bir X-ray dozunda yüksek kaliteli görüntüler sunar. </li><li> Kemik bozulması her iki bacak boyuna bir dakikadan daha kısa bir süre içinde bu sistemi kullanarak izlenebilir. </li><li> Fareler izofluran 3% ile anestezi ve IVIS Spectrum sistemi doğrudan transfer edilebilir. </li><li> İki 60mm görüntüleri tüm hayvan görünümü yakalamak için birleştirilir. </li><li> 30mm FOV görüntüleme yüksek çözünürlüklü diz eklemi için idealdir. </li><li> Kemik metastazı ölçmek ve ölçmek için yeterince yüksek çözünürlükte tek bir tarama sadece 18 saniye sürer ve tek 13mGy bir doz sağlar. </li><li> Fare tümör büyüme ya da hayvan sağlığını etkileyen mikro BT sistemi ile 36 gün içinde 8 kez görüntülenmiştir. </li><li> Mikro BT görüntüleme kemik leisions 10. Gün görünür ve BLI sinyal artışa karşılık, çok hızlı bir şekilde ilerledi gösterdi. </li></ol> 5. BLI ve UCT 3D co-kayıt ve analiz <ol><li> Görüntüleri 3 boyutlu bir yeniden yapılanma içine ortak tescil edilebilir IVIS spektrumu ve Kuantum FX mikro BT sistem veri ele geçirildi. </li><li> MicroCT görüntüler DICOM dosyaları olarak ihraç ve Yaşam Resim 4.1 3D multi modalite aracı yüklenir. </li><li> Yaşayan Görüntü Yazılımı optik sinyal yeniden üç boyutlu DLIT programı çalıştırın. </li><li> Fiducial yazılım aracı, mikro BT görüntüsü optik BLI sinyali co-kayıt için kullanılır. </li><li> Coregistered veri setleri burada, kemik osteolitik lezyon gelişimi ile birlikte, metastatik tümör büyümesini izleme hastalığın ilerlemesi sırasında birden fazla parametre izlemek için kantitatif analiz edilebilir. </li> &lt;/ Ol&gt;

<ol>
	<li>Jenkins, D. E., Hornig, Y. S., Oei, Y., Dusich, J., Purchio, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioluminescent+Human+Breast+Cancer+Cell+Lines+that+Permit+Rapid+and+Sensitive+in+vivo+Detection+of+Mammary+Tumors+and+Multiple+Metastases+in+Immune+Deficient+Mice.">Bioluminescent Human Breast Cancer Cell Lines that Permit Rapid and Sensitive in vivo Detection of Mammary Tumors and Multiple Metastases in Immune Deficient Mice.</a> Breast Cancer Research. 7, R444-R454 (2005).</li><li>Cowey, S., Szafran, A. A., Kappes, J., Zinn, K. R., Siegal, G. P., Desmond, R. A., Kim, H., Evans, L., Hardy, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Breast+Cancer+Metastasis+to+Bone:+Evaluation+of+Bioluminescent+Imaging+and+MicroSPECT/CT+for+Detecting+Bone+Metastasis+in+Immunodeficient+Mice.">Breast Cancer Metastasis to Bone: Evaluation of Bioluminescent Imaging and MicroSPECT/CT for Detecting Bone Metastasis in Immunodeficient Mice.</a> Clinical and Experimental Metastasis. 24, 389-401 (2007).</li><li>Labrinidis, A., Hay, S., Liapis, V., Ponomarev, V., Findlay, D. M., Evdokiou, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zoledronic+Acid+Inhibits+Both+the+Osteolytic+and+Osteoblastic+Components+of+Osteosarcoma+Lesions+in+a+Mouse+Model.">Zoledronic Acid Inhibits Both the Osteolytic and Osteoblastic Components of Osteosarcoma Lesions in a Mouse Model.</a> Clinical Cancer Research. 15, (2009).</li><li>Kaijzel, E. L., Pluijm, G. v. a. n. d. e. r., Lowik, C. W. G. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Whole-Body+Optical+Imaging+in+Animal+Models+to+Assess+Cancer+Development+and+Progression.">Whole-Body Optical Imaging in Animal Models to Assess Cancer Development and Progression.</a> Clinical Cancer Research. 13, 3490-3497 (2007).</li><li>Minn, A. J., Kang, Y., Serganova, I., Gupta, G. P., Giri, D. D., Doubrovin, M., Ponomarev, V., Gerald, W. L., Blasberg, R., Massague, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinct+Organ-Specific+Metastatic+Potential+of+Individual+Breast+Cancer+Cells+and+Primary+Tumors.">Distinct Organ-Specific Metastatic Potential of Individual Breast Cancer Cells and Primary Tumors.</a> The Journal of Clinical Investigation. 115, (2005).</li><li>Xu, H., Christensen, A., Kuo, C., Nilson, D., Lim, E., Whalen, J., Kim, J. B., Zhang, N., Singh, R., Oldfield, S., Troy, T., Rice, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Co-registration+of+in+vivo+Optical+Tomography+and+Micro-CT+for+Longitudinal+Studies.">Co-registration of in vivo Optical Tomography and Micro-CT for Longitudinal Studies.</a> Word Molecular Imaging Congress. , (2010).</li></ol>

Yazarlar bu makalede yer alet kaliper Yaşam Bilimleri çalışanları.

MicroCT Küçük hayvan görüntüleme gibi tümör gelişimi (. Labrinidis ve ark, 2009 Cowey ve ark 2007) gibi patolojik koşullar altında izleme osteolitik lezyonlar tatbik edilmiştir. Lusiferaz etiketli tümörlerin biyoparlaklık görüntüleme yaygın olarak ön-klinik olarak kabul edildiği gibi, biyoparlaklık ve microCT görüntüleri co-kayıt biyoparlaklık sinyalleri (Kaijzel ve ark 2007) anatomik konumu daha iyi bir tanımı. Lusiferaz etiketli bir meme tümörü hücre hattı MDA-MB-231-luc-D3H2LN Önceki çalışmada kalp içi enjeksiyonu takiben kemik ve osteolitik lezyonların gelişimi metastaz gösterdi (Jenkins ve ark 2005; Minn ve ark, 2005). Bu çalışmada, sistemik MDA-MB-231-luc-D3H2LN metastaz modeli ile combinatory optik ve düşük radyasyon dozu microCT görüntüleme ile tümörün metastaz ve osteolitik aktivite uzunlamasına bir çalışmada araştırdı. Biz IVIS Spectrum ve Kuantum FX microCT sistemleri ve elde biyoparlaklık ve CT ile birlikte yeni geliştirilen Oturma Image 4.1 yazılımı (Heng ve ark, 2010) kaydı ile sıralı görüntüleme yapıldı. IVIS ile Biyoparlaklık görüntüleme tümörler erken aşamada tespit edilmesini sağlayarak, yüksek hassasiyet ile tümör hücrelerinin non-invaziv algılanmasını sağlar. Kuantum FX ile görüntüleme, kemik lezyonları ve detaylı anatomik bilgi doğru 3 boyutlu rekonstrüksiyon sağlar. Kuantum döviz sistemi, hastalığın ilerlemesi boyuna izlenmesini sağlar düşük radyasyon dozu görüntüleme sunar. Kemik lezyonları ve tümör metastazı arasındaki korelasyon eş kayıtlı görüntü tasvir edildi ve lezyon gelişimi multimodalite izleme avantajı boyuna açıkça göstermiştir.

monitoring tumor metastases and osteolytic lesions with bioluminescence and micro ct imaging

Nu / Nu fareler MDA-MB-231-luc-D3H2LN hücrelerinin sonra intrakardiyak teslimat, enjekte edilen hayvanların geliştirilen sistemik metastaz. Biyoparlaklık görüntüleme IVIS Spektrum ile dağıtım ve tümör sinyal ve yerini ölçmek için DLIT yeniden inşası da dahil olmak üzere dağıtım prosedürü izleyerek tümör hücrelerinin gelişimini izlemek için istihdam edilmiştir. Metastatik lezyonlar kemik dokulara Geliştirme tibia osteolitik aktivite ve lezyonlar tetikler ve femur mikro BT kullanarak boyuna değerlendirildi. Görüntüleme, hızlı görüntüleme ve düşük X-ray dozu ile bir Kuantum FX mikro BT sistemi kullanılarak yapıldı. Düşük radyasyon dozu, çoklu görüntüleme oturumları çok LD50 altında kümülatif bir X-ışını doz ile yapılacak sağlar. Bir fare görüntüleme servis cihazı görüntü sırayla IVIS Spectrum ve Kuantum FX ile fareler biyoparlaklık ve BT görüntüleri hem de doğru hayvan konumlandırma elde kullanılmıştır. Yaşayan Image 4.1 yazılımı kullanarak optik ve BT veri setleri 3-Boyutları, eş-tescil edilmiştir. Bu multi-mode yaklaşım, yakın izleme osteolitik faaliyet ile eş zamanlı olarak tümör büyüme ve gelişme sağlar.

Watch this Scientific Journal Video about Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging at JoVE.com

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

Kemik metastazı deneysel bir fare modeli lusiferaz meme tümör hücreleri ifade intrakardiyak teslim edildikten sonra kurulmuştur. Biyoparlaklık ve mikro BT görüntüleme ile tümör gelişimi ve osteolitik lezyon sonuçlandı boyuna takip edildi.

Biyoparlaklık ve Micro CT Görüntüleme Tümör Metastazlari ve osteolitik Lezyonlar İzleme

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

monitoring-tumor-metastases-osteolytic-lesions-with-bioluminescence

Nanyang Technological University

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Medicine

24.5K Görüntüleme.  Caliper Life Sciences.  中文 

Video: Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

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.

Meme Kanseri Beyin metastaz tümör Hipoksi Dynamics in vivo Biyoparlaklık Görüntüleme Fare Modeli

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

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A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a novel platform that allows for direct long-term visualization of tissue structure changes intracranially. Imaging at a single cell resolution in a real-time fashion provides supplementary dynamic information beyond that provided by standard end-point histological analysis, which looks solely at 'snap-shot' cross sections of tissue.
Establishing this intravital imaging technique in fluorescent chimeric mice, we are able to image four fluorescent channels simultaneously. By incorporating fluorescently labeled cells, such as GFP+ bone marrow, it is possible to track the fate of these cells studying their long-term migration, integration and differentiation within tissue. Further integration of a secondary reporter cell, such as an mCherry glioma tumor line, allows for characterization of cell:cell interactions. Structural changes in the tissue microenvironment can be highlighted through the addition of intra-vital dyes and antibodies, for example CD31 tagged antibodies and Dextran molecules.
Moreover, we describe the combination of our ICW imaging model with a small animal micro-irradiator that provides stereotactic irradiation, creating a platform through which the dynamic tissue changes that occur following the administration of ionizing irradiation can be assessed.
Current limitations of our model include penetrance of the microscope, which is limited to a depth of up to 900 &mu;m from the sub cortical surface, limiting imaging to the dorsal axis of the brain. The presence of the skull bone makes the ICW a more challenging technical procedure, compared to the more established and utilized chamber models currently used to study mammary tissue and fat pads 5-7. In addition, the ICW provides many challenges when optimizing the imaging.

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

We describe a novel in vivo imaging technique that couples fluorescent chimeric mice with intracranial windows and high-resolution 2-photon microscopy. This imaging platform aids studies of dynamic changes in brain tissue and microvasculature, at a single-cell level, following pathological insults and is adaptable to assess intracranial drug delivery and distribution.

Yeni Bir Yüksek çözünürlüklü<em&gt; In vivo</emTümör Büyüme ve Tedavi için İntrakranial Yapıların Dinamik Tepki Eğitim&gt; Görüntüleme Tekniği

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to stimulate microcirculation. The choice of delivery method for these agents is important because the route of administration profoundly affects the bioactivity and efficacy of these agents1,2. In this article, we demonstrate how to locally administer a substance in ischemic hindlimbs by using a catheterized osmotic pump. This pump can deliver a fixed volume of aqueous solution continuously for an allotted period of time. We also present our mouse model of unilateral hindlimb ischemia induced by ligation of the common femoral artery proximal to the origin of profunda femoris and epigastrica arteries in the left hindlimb. Lastly, we describe the in vivo cannulation and ligation of the infrarenal abdominal aorta and perfusion of the hindlimb vasculature with Microfil, a silicone radiopaque casting agent. Microfil can perfuse and fill the entire vascular bed (arterial and venous), and because we have ligated the major vascular conduit for exit, the agent can be retained in the vasculature for future ex vivo imaging with the use of small specimen micro-CT3.

We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.

Mikro-Bilgisayarlı Tomografi Görüntüleme için Microfil ile İskemik arka ayaklarında ve damarları bir Perfüzyon için osmotik İlaç Taşıyıcı

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Bioluminescence Imaging of an Immunocompetent Animal Model for Glioblastoma

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and histopathologic features of the human disease. When GL261 cells are implanted intracranially in syngeneic C57BL/6 mice, the model has the added advantage of maintaining an intact immune microenvironment. Stable expression of luciferase in GL261 cells allows non-invasive cost effective bioluminescence monitoring of intracranial tumor growth. We have recently demonstrated that luciferase expression in GL261 cells does not affect the tumor growth properties, tumor cell immunomodulatory cytokine expression, infiltration of immune cells into the tumor, or overall survival of animals bearing the intracranial tumor. Therefore, it appears that the GL261 luciferase glioma model can be useful in the study of novel chemotherapeutic and immunotherapeutic modalities. Here we report the technique for generating stable luciferase expression in GL261 cells and how to study the in vitro and in vivo growth of the tumor cells by bioluminescence imaging.

GL261 glioma cells provide a useful immunocompetent animal model of glioblastoma. The goals of this protocol are to demonstrate proper techniques for monitoring intracranial tumor growth using in vivo bioluminescence imaging, and to verify the utility of luciferase-modified GL261 cells for studying tumor immunology and immunotherapeutic approaches for treating glioblastoma.

Glioblastoma&#39;nın için bir Immünkompetan Hayvan Modeli Biyoparlaklık Görüntüleme

In contrast to commonly reported human glioma xenograft animal models, GL261 murine glioma xenografts recapitulate nearly all relevant clinical and ...

Bioluminescence-Based Tumor Quantification Method for Monitoring Tumor Progression and Treatment Effects in Mouse Lymphoma Models

Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. Here we describe a method for bioluminescence imaging to obtain a semi-quantitative analysis of tumor burden and treatment response. This method is based on the calculation of a luminoscore, a value that allows comparisons of two animals from the same or different experiments. Current BLI instruments enable the calculation of this luminoscore, which relies mainly on the acquisition conditions (back and front acquisitions) and the drawing of the region of interest (manual markup around the mouse). Using two previously described mouse lymphoma models based on cell engraftment, we show that the luminoscore method can serve as a noninvasive way to verify successful tumor cell inoculation, monitor tumor burden, and evaluate the effects of in situ cancer treatment (CpG-DNA). Finally, we show that this method suits different experimental designs. We suggest that this method be used for early estimates of treatment response in preclinical small-animal studies.

Bioluminescence imaging is a well-known tool for localizing tumors and metastases, but quantification of these images often requires complex calculations and particular instruments. We describe the easy-to-use luminoscore method, based on precise acquisition conditions, requiring no calculations, and enabling tumor burden and treatment response to be monitored in mouse models.

Fare Lenfoma Modellerinde İzleme Tümör ilerleme ve Tedavi Etkileri Biyoparlaklık Tabanlı Tümör Niceleme Yöntemi

Although bioluminescence imaging (BLI) shows promise for monitoring tumor burden in animal models of cancer, these analyses remain mostly qualitative. ...

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of stroke than relying on indirect measurements, and will be a potent and robust vascular research tool. We use an optical imaging approach that labels thrombi with a molecular imaging thrombus marker&#160;&#8212; a Cy5.5 near-infrared fluorescent (NIRF) probe that is covalently linked to the fibrin strands of the thrombus by the fibrin-crosslinking enzymatic action of activated coagulation factor XIIIa during the process of clot maturation. A micro-computed tomography (microCT)-based approach uses thrombus-seeking gold nanoparticles (AuNPs) functionalized to target the major component of the clot: fibrin. This paper describes a detailed protocol for the combined in vivo microCT and ex vivo NIRF imaging of thromboemboli in a mouse model of embolic stroke. We show that in vivo microCT and fibrin-targeted glycol-chitosan AuNPs (fib-GC-AuNPs) can be used for visualizing both in situ thrombi and cerebral embolic thrombi. We also describe the use of in vivo microCT-based direct thrombus imaging to serially monitor the therapeutic effects of tissue plasminogen activator-mediated thrombolysis. After the last imaging session, we demonstrate by ex vivo NIRF imaging the extent and the distribution of residual thromboemboli in the brain. Finally, we describe quantitative image analyses of microCT and NIRF imaging data. The combined technique of direct thrombus imaging allows two independent methods of thrombus visualization to be compared: the area of thrombus-related fluorescent signal on ex vivo NIRF imaging vs. the volume of hyperdense microCT thrombi in vivo.

This protocol describes the application of combined near-infrared fluorescent (NIRF) imaging and micro-computed tomography (microCT) for visualizing cerebral thromboemboli. This technique allows the quantification of thrombus burden and evolution. The NIRF imaging technique visualizes fluorescently labeled thrombus in excised brain, while the microCT technique visualizes thrombus inside living animals using gold-nanoparticles.

Doğrudan Serebral emboli görselleştirme için Kombine Yakın kızılötesi Floresan Görüntüleme ve Mikro bilgisayarlı Tomografi

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

Kullanımı Glukoz Metabolizma Kinetiği Belirlenmesi<sup&gt; 18</sup&gt; F-FDG Mikro PET / BT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing motor function deficits are basic readouts of the immune-mediated inflammation of the spinal cord. However, scores and body weight do not allow for an in vivo assessment of brain inflammation and optic neuritis. The latter is an early and frequent manifestation in about 2/3 of MS patients. Here, we show methods for bioluminescence and near-infrared live imaging to assess EAE evoked optic neuritis, brain inflammation, and blood-brain barrier (BBB) disruption in living mice using an in vivo imaging system. A bioluminescent substrate activated by oxidases primarily showed optic neuritis. The signal was specific and allowed the visualization of medication effects and disease time courses, which paralleled the clinical scores. Pegylated fluorescent nanoparticles that remained within the vasculature for extended periods of time were used to assess the BBB integrity. Near-infrared imaging revealed a BBB leak at the peak of the disease. The signal was the strongest around the eyes. A near-infrared substrate for matrix metalloproteinases was used to assess EAE-evoked inflammation. Auto-fluorescence interfered with the signal, requiring spectral unmixing for quantification. Overall, bioluminescence imaging was a reliable method to assess EAE-associated optic neuritis and medication effects and was superior to the near-infrared techniques in terms of signal specificity, robustness, ease of quantification, and cost.

We show a technique for in vivo live bioluminescence and near-infrared imaging of optic neuritis and encephalitis in the experimental autoimmune encephalomyelitis (EAE) model for multiple sclerosis in SJL/J mice.

Farelerde Multipl Skleroz EAE Modeli Optik Nörit ve Beyin Enflamasyon Biyoparlaklık ve Yakın-Kızılötesi Görüntüleme

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing ...

Induction and Micro-CT Imaging of Cerebral Cavernous Malformations in Mouse Model

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have been established by tamoxifen induced deletion of Ccm genes in postnatal animals. These mouse models provide invaluable tools to investigate molecular mechanism and therapeutic approaches for CCM disease. An accurate and quantitative method to assess lesion burden and progression is essential to harness the full value of these animal models. Here, we demonstrate the induction of CCM disease in a mouse model and the use of the contrast enhanced X-ray micro computed tomography (micro-CT) method to measure CCM lesion burden in mouse brains. At postnatal day 1 (P1), we used 4-hydroxytamoxifen (4HT) to activate Cre recombinase activity from the Cdh5-CreErt2 transgene to cleave the floxed allele of Ccm2. CCM lesions in mouse brains were analyzed at P8. For micro-CT, iodine based Lugol&#39;s solution was used to enhance contrast in brain tissue. We have optimized the scan parameters and utilized a voxel dimension of 9.5 &#181;m, which lead to a minimum feature size of approximately 25 &#181;m. This resolution is sufficient to measure CCM lesion volume and number globally and accurately, and provide high-quality 3-D mapping of CCM lesions in mouse brains. This method enhances the value of the established mouse models to study the molecular basis and potential therapies for CCM and other cerebrovascular diseases.

This protocol demonstrates the induction of cerebral cavernous malformation disease in a mouse model and uses contrast enhanced micro computed tomography to measure lesion burden. This method enhances the value of established mouse models to study the molecular basis and potential therapies for cerebral cavernous malformation and other cerebrovascular diseases.

İndüksiyon ve fare modeli serebral derin malformasyon mikro-CT görüntüleme

Mutations in the CCM1 (aka KRIT1), CCM2, or CCM3 (aka PDCD10) gene cause cerebral cavernous malformation (CCM) in humans. Mouse models of CCM disease have ...

Gene Regulation and Targeted Therapy in Gastric Cancer Peritoneal Metastasis: Radiological Findings from Dual Energy CT and PET/CT

Gastric cancer remains fourth in cancer incidence worldwide with a five-year survival of only 20%-30%. Peritoneal metastasis is the most frequent type of metastasis that accompanies unresectable gastric cancer and is a definitive determinant of prognosis. Preventing and controlling the development of peritoneal metastasis could play a role in helping to prolong the survival of gastric cancer patients. A non-invasive and efficient imaging technique will help us to identify the invasion and metastasis process of peritoneal metastasis and to monitor the changes in tumor nodules in response to treatments. This will enable us to obtain an accurate description of the development process and molecular mechanisms of gastric cancer. We have recently described experiment using dual energy CT (DECT) and positron emission tomography/computed tomography (PET/CT) platforms for the detection and monitoring of gastric tumor metastasis in nude mice models. We have shown that weekly continuous monitoring with DECT and PET/CT can identify dynamic changes in peritoneal metastasis. The sFRP1-overexpression in gastric cancer mice models showed positive radiological performance, a higher FDG uptake and increasing enhancement, and the SUVmax (standardized uptake value) of nodules demonstrated an obvious alteration trend in response to targeted therapy of TGF-&#946;1 inhibitor. In this article, we described the detailed non-invasive imaging procedures to conduct more complex research on gastric cancer peritoneal metastasis using animal models and provided representative imaging results. The use of non-invasive imaging techniques should enable us to better understand the mechanisms of tumorigenesis, monitor tumor growth, and evaluate the effect of therapeutic interventions for gastric cancer.

This protocol describes the value of dual energy CT and PET/CT imaging methods in tumor imaging and efficacy evaluation. This article demonstrates the research methods and results acquired by dual energy CT and PET/CT to evaluate the gene regulation and targeted treatment of gastric cancer peritoneal metastasis.

Gen düzenlemesi ve hedefe yönelik tedavi mide kanseri Periton metastaz: Dual Enerji CT ve PET/BT radyolojik bulgular

Gastric cancer remains fourth in cancer incidence worldwide with a five-year survival of only 20%-30%. Peritoneal metastasis is the most frequent type of ...