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

Guided Tumor Inoculation and Multimodal Imaging in Mice

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.

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

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

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.

종양 성장 시각화 및 이미징을 위한 종양 세포 주입 마우스의 창 챔버 수술

시작하려면 메인 창 챔버 부품을 높은 수준의 액체 소독제로 12분 동안 살균합니다. 그런 다음 이소프로필 알코올에 5분 동안 담근다. 종양 세포가 주입된 마우스에게 수술 1-3시간 전에 서방형 부프레노르핀을 킬로그램당 1mg씩 투여합니다.소독을 위해 마취하고 면도한 쥐를 7.5% 포비돈-요오드 용액으로 문지릅니다. 70% 이소프로필 알코올로 소독 스크럽을 닦아냅니다. 10% 포비돈-요오드 용액을 피부에 바릅니다.마지막으로 수의학 눈 윤활제를 바르고 건조함을 방지합니다. 척추를 따라 쥐의 피부를 들어 올리고 측면 피부 주름 중 하나에서 자라는 종양을 찾습니다. 등쪽 피부접힘 창실(DSFC)의 뒤쪽 프레임을 배치하여 종양이 프레임 내 중앙에 오도록 합니다.DSFC의 상단 3개 구멍에 3개의 봉합사를 추가하고 수술 가이드를 추가하여 백 프레임의 위치를 고정합니다. DSFC의 후면 프레임을 통해 스페이서가 들어 있는 구멍에 세 개의 바늘을 삽입합니다. 수술용 마커를 사용하여 피부의 DSFC 백 프레임 스페이서에 대한 세 점을 표시합니다.제거할 피부 부위를 나타내기 위해 지름 1cm의 원을 그립니다. 세 개의 점을 추가로 표시한 후 봉합사를 잘라 바늘과 뒷부분을 제거합니다. 수술용 마이크로 가위를 사용하여 표시된 원에서 피부를 제거합니다.그런 다음 임시 봉합사를 배치하여 창실 부분을 피부에 고정합니다. 임시 봉합사로 어셈블리를 조이고 구멍을 통해 피부 아래에 전면 프레임을 삽입합니다. DSFC의 전면 및 후면 프레임 둘레에 있는 구멍을 통해 영구 봉합사를 배치합니다.임시 봉합사를 제거한 후 지지 클립을 DSFC 백 프레임에 부착합니다. 통증과 염증 감소를 위해 체중 1kg당 5mg의 멜록시캠을 마우스에 피하로 투여합니다.

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 ...

연구

암 연구학

Tumor Cells Inoculation in Mice Through Precise Landmarking and Injection Techniques

정밀한 랜드마크 및 주입 기술을 통한 생쥐의 종양 세포 접종

Integrative Imaging Techniques for Tumor Analysis in Mice

생쥐의 종양 분석을 위한 통합 이미징 기법