Abstract
Intravital imaging is an invaluable tool for studying the expanding range of immune cell functions. Only in vivo can the complex and dynamic behavior of leukocytes and their interactions with their natural microenvironment be observed and quantified. While the capabilities of high-speed, high-resolution confocal and multiphoton microscopes are well-documented and steadily improving, other crucial hardware required for intravital imaging is often developed in-house and less commonly published in detail. In this report, we describe a low-cost, multipurpose, and tissue-stabilizing in vivo imaging platform that enables sensing and regulation of local tissue temperature. The effect of tissue temperature on local blood flow and leukocyte migration is demonstrated in muscle and skin. Two different models of vacuum windows are described in this report, however, the design of the vacuum window can easily be adapted to fit different organs and tissues.
Highlights
IntroductionMore advanced microscope technologies have been steadily emerging, providing researchers with a plethora of options for visualization of complex biological processes
Microscopy has a long history as an important tool in life science research
The purpose of the vacuum window imaging platform is to immobilize tissue from an anesthetized animal to enable in vivo high resolution imaging, primarily using an upright confocal or multi photon microscope
Summary
More advanced microscope technologies have been steadily emerging, providing researchers with a plethora of options for visualization of complex biological processes. With the invention of the confocal microscope [1] and its ability to optically section the imaged tissue, the possibility of imaging cell-cell interactions in vivo was greatly improved. Further technological advances allowed for the introduction of the multiphoton microscope, which improved some of the limiting aspects of confocal microscopy, such as phototoxicity and penetration depth. While these microscopy techniques enable high temporal and spatial resolution imaging, other aspects of in vivo imaging remain challenging, and impact the quality of the results. Devices using vacuum for immobilization of the lungs and other organs have been previously described, but has required custom-made metallic parts to function, somewhat increasing the threshold for most researchers to acquire such systems [2,3,4,5]
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