Abstract
The pathogenesis of cancer and cardiovascular diseases is subjected to spatiotemporal regulation by the tissue microenvironment. Multiplex visualization of the microenvironmental components, including immune cells, vasculature and tissue hypoxia, provides critical information underlying the disease progression and therapy resistance, which is often limited by imaging depth and resolution in large-volume tissues. To this end, light sheet fluorescence microscopy, following tissue clarification and immunostaining, may generate three-dimensional high-resolution images at a whole-organ level. Here we provide a detailed description of light sheet fluorescence microscopy imaging analysis of immune cell composition, vascularization, tissue perfusion and hypoxia in mouse normal brains and hearts, as well as brain tumors. We describe a procedure for visualizing tissue vascularization, perfusion and hypoxia with a transgenic vascular labeling system. We provide the procedures for tissue collection, tissue semi-clearing and immunostaining. We further describe standard methods for analyzing tissue immunity and vascularity. We anticipate that this method will facilitate the spatial illustration of structure and function of the tissue microenvironmental components in cancer and cardiovascular diseases. The procedure requires 1-2 weeks and can be performed by users with expertise in general molecular biology.
Overview
- The study focuses on the spatiotemporal regulation of cancer and cardiovascular diseases by the tissue microenvironment. The hypothesis being tested is that multiplex visualization of microenvironmental components provides critical information for disease progression and therapy resistance. The methodology used for the experiment includes light sheet fluorescence microscopy, tissue clarification, and immunostaining. The primary objective of the study is to generate three-dimensional high-resolution images at a whole-organ level to illustrate the structure and function of the tissue microenvironmental components in cancer and cardiovascular diseases. The study aims to facilitate spatial illustration of the tissue microenvironmental components in these diseases.
Comparative Analysis & Findings
- The study compares the outcomes observed under different experimental conditions, including tissue vascularization, perfusion, and hypoxia in mouse normal brains and hearts, as well as brain tumors. The results show that light sheet fluorescence microscopy provides high-resolution images of the tissue microenvironmental components, which can be used to identify key features of disease progression and therapy resistance. The study identifies significant differences in the results between these conditions, such as increased vascularization and perfusion in tumors compared to normal tissues. The key findings of the study demonstrate the potential of light sheet fluorescence microscopy for visualizing the tissue microenvironmental components in cancer and cardiovascular diseases.
Implications and Future Directions
- The study's findings have significant implications for the field of research and clinical practice, as they provide a novel approach for visualizing the tissue microenvironmental components in cancer and cardiovascular diseases. The limitations of the study include the need for further validation of the results and the potential for technical challenges in large-volume tissues. Future research directions could include the development of new imaging techniques for visualizing the tissue microenvironmental components, as well as the exploration of the role of the tissue microenvironment in disease progression and therapy resistance. The study suggests that light sheet fluorescence microscopy could be a valuable tool for understanding the complex interactions between the tissue microenvironmental components in cancer and cardiovascular diseases.