window chamber surgery in tumor cell injected mice for tumor growth visualization and imaging

चूहों में निर्देशित ट्यूमर टीकाकरण और मल्टीमॉडल इमेजिंग

Tumor microvasculature is an important component of the tumor microenvironment that can be a target for therapy and a determinant of treatment response. In the preclinical setting, the microvasculature is typically studied using intravital microscopy in orthotopic or heterotopic window chamber animal models1,2. This has several advantages over histological studies since the imaging is done in live tissues and the tumor can be monitored longitudinally over several weeks or even months2,3. These studies can leverage the high-resolution imaging capabilities of intravital microscopy to study the delivery of therapeutics to the tumor4,5, the causes of treatment resistance6, and the response of the micro vessels to therapies such as antiangiogenic treatment7,8 and radiotherapy2,9.
Intravital microscopy clearly plays an important role in preclinical cancer research; however, how can tumor microenvironmental features be measured in the clinic? Microvascular information would be useful in the clinic for measuring blood supply and tumor cell hypoxia, which is important for determining treatment resistance in radiotherapy10, as well as the ability of the microvasculature to deliver chemotherapeutic agents to the surrounding tumor cells11. For example, in radiotherapy, spatial information on the structure and function of the tumor microvasculature may help personalize a patient&#39;s treatment plan by adjusting the fractionation schedule or by preferentially boosting the dose to avascular and likely hypoxic regions12.
Intravital microscopy can measure these important microvascular features since it has a very high resolution (&#956;m scale); however, its depth penetration into tissue is limited to several hundred microns or a few millimeters, at most making clinical implementation challenging. Indeed, there are some novel applications of intravital microscopy in the clinic13; however, these are still limited to examinations of near-surface level tissue such as the skin14 or mucosal/endothelial linings of various body cavities via flexible catheters/endoscopes15,16.
More commonly, the microvasculature is studied using imaging modalities such as CT17 or MRI18. These clinical imaging modalities can image to any depth within the body, but they have a much lower spatial resolution (mm scale). Thus, there is a need to bridge this resolution gap between preclinical intravital microscopy and clinical imaging modalities to bring high-resolution and detailed microvascular information into the clinic19. Several functional imaging methods have been developed to improve the microvascular imaging capabilities of clinical imaging modalities such as dynamic contrast-enhanced (DCE) MRI and CT20, and Intravoxel incoherent motion (IVIM) MRI21. However, these are model-based methods that provide indirect measurements of the microvasculature and thus, must be validated with appropriate &#34;ground truth&#34; measurements of the microvasculature19,22.
We have developed a dorsal skinfold window chamber (DSFC) tumor mouse model to bridge this gap between preclinical intravital microscopy and clinically applicable imaging modalities such as CT and MRI. The DSFC provides direct access to the tumor for high-resolution, intravital microscopy imaging through a glass window but also clinically applicable imaging such as MRI as it is made of MR-compatible materials (plastic and glass). Furthermore, an included MATLAB code performs multimodality 3D co-registration for direct spatial correlations between preclinical intravital microscopy and clinically applicable imaging modalities. Here we will describe the design and surgery to install the DSFC as well as the procedure to co-register intravital microscopy and clinically applicable imaging modalities.

लेखक स्पॉटलाइट: स्टीरियोटैक्टिक बॉडी रेडिएशन थेरेपी प्लानिंग को बढ़ाने के लिए उच्च-रिज़ॉल्यूशन इंट्रावाइटल इमेजिंग और एमआरआई को एकीकृत करना

ट्रांसलेशनल कैंसर रिसर्च में संयुक्त इंट्रावाइटल माइक्रोस्कोपी और चुंबकीय अनुनाद इमेजिंग के लिए एक पृष्ठीय स्किनफोल्ड विंडो चैंबर ट्यूमर माउस मॉडल

Preclinical intravital imaging has very high resolution, but limited depth penetration in the tissue, which makes it very good for preclinical imaging studies. On the other hand, MRI is much more clinically applicable and has a higher depth penetration, but very low spatial resolution. The goal of this study is to correlate these two modalities with each other in order to better translate our findings from preclinical intravital microscopy into the clinic via magnetic resonance imaging.Perfusion-sensitive imaging methods in MRI and biophotonics, including our optical coherence tomography and geography system, are revealing how the microvasculature impacts tumor response to hypofractionated radiotherapy. 3D printing is also aiding through cost-effective custom tool manufacturing, facilitating longitudinal studies. Finally, AI image analysis is facilitating the identification of novel radiotherapy predictive biomarkers.We've made several findings concerning the tumor response to hypofractionated radiotherapy, including more recently, the temporal kinetics of both radiobiologically significant and abstracted AI-derived tissue response metrics. Additionally, we have demonstrated the promise for multimodal imaging being performed more accurately, correlating both MRI, macroscopic and optical coherence tomography, microscopic, angiography-derived metrics. By extending the effective length of longitudinal studies, we may better identify predictive biomarkers for long-term tumor treatment response.Additionally, by improving robustness of co-registration between macroscopic clinically available imaging modalities and microscopic preclinical imaging modalities, we may enhance the translatability of our findings from benchtop to bedside. During a stereotactic body radiation therapy, high doses of radiation are delivering a reduced number of fractions. One potential radiobiological target associated with increased cell death in SVRT is the tumor microvasculature.By combining high-resolution optical angiography and MRI, we want to enable functional MRI microvascular imaging for patient-specific SVRT treatment planning and improved outcomes.

ट्यूमर सेल में विंडो चैंबर सर्जरी ट्यूमर ग्रोथ विज़ुअलाइज़ेशन और इमेजिंग के लिए इंजेक्शन चूहों

शुरू करने के लिए, 12 मिनट के लिए एक उच्च स्तरीय तरल कीटाणुनाशक में मुख्य खिड़की कक्ष भागों बाँझ. पांच मिनट के लिए आइसोप्रोपिल अल्कोहल में भिगोने के बाद। सर्जरी से एक से तीन घंटे पहले ट्यूमर सेल इंजेक्शन माउस के लिए निरंतर रिलीज buprenorphine के किलोग्राम प्रति मिलीग्राम प्रशासन.कीटाणुशोधन के लिए, 7.5% पोविडोन-आयोडीन समाधान के साथ संवेदनाहारी और मुंडा माउस को साफ़ करें। 70% आइसोप्रोपिल अल्कोहल के साथ कीटाणुनाशक स्क्रब को साफ़ करें। त्वचा पर 10% पोविडोन-आयोडीन समाधान लागू करें।अंत में, सूखापन को रोकने के लिए पशु चिकित्सा आंख स्नेहक लागू करें। रीढ़ की हड्डी के साथ माउस की त्वचा लिफ्ट और पार्श्व skinfolds में से एक पर बढ़ ट्यूमर का पता लगाएं. पृष्ठीय स्किनफोल्ड विंडो चैंबर, या डीएसएफसी के पीछे के फ्रेम की स्थिति बनाएं, ताकि ट्यूमर फ्रेम के भीतर केंद्रित हो।डीएसएफसी के शीर्ष तीन छेदों में तीन टांके जोड़ें और पीछे के फ्रेम की स्थिति को ठीक करने के लिए सर्जिकल गाइड। स्पेसर युक्त छेद में DSFC के पीछे फ्रेम के माध्यम से तीन सुइयों डालें. सर्जिकल मार्कर का उपयोग करके, त्वचा पर डीएसएफसी बैक फ्रेम स्पेसर्स के लिए तीन बिंदुओं को चिह्नित करें।हटाए जाने वाले त्वचा क्षेत्र को इंगित करने के लिए एक सेंटीमीटर व्यास सर्कल बनाएं। तीन अतिरिक्त डॉट्स को चिह्नित करने के बाद, सुइयों और बैक फ्रेम को हटाने के लिए टांके काट लें। सर्जिकल माइक्रो कैंची का उपयोग करके चिह्नित सर्कल से त्वचा निकालें।फिर त्वचा के लिए खिड़की कक्ष भागों सुरक्षित करने के लिए अस्थायी सिवनी जगह. एक अस्थायी सिवनी के साथ विधानसभा को कस लें और छेद के माध्यम से त्वचा के नीचे सामने के फ्रेम को डालें। डीएसएफसी के सामने और पीछे के फ्रेम की परिधि के आसपास छेद के माध्यम से स्थायी टांके लगाएं।अस्थायी सिवनी को हटाने के बाद, समर्थन क्लिप को DSFC बैक फ्रेम में चिपका दें। दर्द और सूजन में कमी के लिए माउस को चमड़े के नीचे प्रति किलोग्राम मेलॉक्सिकैम के पांच मिलीग्राम प्रशासित करें।

window-chamber-surgery-tumor-cell-injected-mice-for-tumor-growth

Research

JoVE Journal

Cancer Research

Window Chamber Surgery in Tumor Cell Injected Mice for Tumor Growth Visualization and Imaging

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 ...