Abstract
In vivo, longitudinal observation of tumorigenesis in a live mouse model over an extended time period has been actively pursued to obtain a better understanding of the cellular and molecular mechanism in a highly complex tumor microenvironment. However, common intravital imaging approaches based on a conventional laser scanning confocal or a two-photon microscope have been mostly limited to the observation of superficial parts of the solid tumor tissue. In this work, we implemented a small diameter needle-shaped side-view confocal endomicroscope that can be directly inserted into a solid tumor in a minimally-invasive manner in vivo. By inserting the side-view endomicroscope into the breast tumor from the surface, we achieved in vivo depth-wise cellular-level visualization of microvasculature and fluorescently labeled tumor cells located deeply inside the tumor. In addition, we successfully performed longitudinal depth-wise visualization of a growing breast tumor over three weeks in a live mouse model, which revealed dynamic changes in microvasculature such as a decreasing amount of intratumoral blood vessels over time.
Highlights
Tumor microenvironments are constituted by tumor cells and numerous stromal cells and immune cells with blood vessels and various soluble factors such as hypoxiainducible factors
A longitudinal in vivo cellular-level visualization of deep tumor tissue during tumorigenesis to investigate the dynamically changing microenvironment and complex pathophysiology has remained as a challenging task
2.1 Needle-shaped side-view endomicroscope based on GRIN lens
Summary
Tumor microenvironments are constituted by tumor cells and numerous stromal cells and immune cells with blood vessels and various soluble factors such as hypoxiainducible factors. Highresolution intravital imaging has been mostly limited to the superficial portion of the tumor tissues This is mainly due to a rapidly deteriorating signal-to-background ratio in the image acquired at a deeper portion of tissues, which is mainly caused by increased multiple light scattering. To overcome this limitation, a miniature optical probe based on graded-index (GRIN) rod lenses was suggested to enable cellular-level deep tissue imaging [22,23,24,25,26,27,28,29]. A longitudinal in vivo cellular-level visualization of deep tumor tissue during tumorigenesis to investigate the dynamically changing microenvironment and complex pathophysiology has remained as a challenging task
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