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Updated: Jun 12, 2025

Longitudinal Intravital Imaging Through Clear Silicone Windows
Published on: January 5, 2022
Longitudinal intravital microscopy of the mouse kidney: inflammatory responses to abdominal imaging windows
Michelle M Martinez1, Julia R Walsh2, Malgorzata M Kamocka1
1Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana, United States.
Abstract:
Intravital microscopy enables direct observation of cell biology and physiology at subcellular resolution in real time in living animals. Implanted windows extend the scope of intravital microscopy to processes extending for weeks or even months, such as disease progression or tumor development. However, a question that must be addressed in such studies is whether the imaging window, like any foreign body, triggers an inflammatory response, and whether that response alters the biological process under investigation. To directly evaluate this question, we conducted large-scale intravital microscopy of the kidney of LysM-EGFP mice over time after implantation of abdominal imaging windows. These studies demonstrate that windows stimulated a variety of changes consistent with a foreign body response. Within a few days of implantation, leukocytes were recruited to the window and the region between the window and kidney where, over the next 16 days, they increased in number in an expanding volume that developed a new vascular network. These changes were accompanied by a dramatic increase in glomerular albumin permeability within 2-5 days of implantation. Similar results were obtained from mice implanted with windows coated with poly(l-lysine)-graft-polyethylene glycol (PLL-g-PEG), but not from immune-deficient mice. These studies demonstrate the importance of evaluating whether implanted windows induce an inflammatory response, and whether that response impacts the processes under evaluation in longitudinal intravital microscopy studies.NEW & NOTEWORTHY Intravital microscopy studies of LysM-EGFP mice demonstrate that abdominal imaging windows placed over the kidney stimulated a variety of changes consistent with a foreign body response. Within a day of implantation, leukocytes were recruited to the window where, over the next 16 days, they increased in number in an expanding volume that developed a new vascular network. These changes were accompanied by a dramatic increase in glomerular permeability to albumin.
Insights
Implanted imaging windows for intravital microscopy can trigger a foreign body response in mice, recruiting leukocytes and increasing kidney glomerular permeability. This inflammation may affect long-term studies of biological processes.
Area of Science:
- Biological Sciences
- Medical Imaging
- Immunology
Background:
- Intravital microscopy allows real-time, subcellular observation of biological processes in vivo.
- Implanted windows enable long-term intravital microscopy studies, but their potential to elicit foreign body responses is a concern.
Purpose of the Study:
- To investigate whether abdominal imaging windows used in intravital microscopy trigger an inflammatory response.
- To determine if this inflammatory response impacts biological processes, specifically kidney function.
Main Methods:
- Large-scale intravital microscopy was performed on LysM-EGFP mice over time after abdominal imaging window implantation.
- Recruitment and proliferation of leukocytes, development of new vasculature, and changes in glomerular albumin permeability were monitored.
- Studies included windows coated with poly(l-lysine)-graft-polyethylene glycol (PLL-g-PEG) and immune-deficient mice.
Main Results:
- Implanted windows induced a foreign body response, characterized by leukocyte recruitment within days.
- An expanding volume of leukocytes developed a new vascular network over 16 days post-implantation.
- A significant increase in glomerular albumin permeability was observed within 2-5 days, indicating altered kidney function.
Conclusions:
- Abdominal imaging windows can elicit a foreign body response in mice, involving leukocyte infiltration and neovascularization.
- This inflammatory response leads to increased glomerular permeability, potentially confounding long-term intravital microscopy studies.
- Researchers must evaluate the impact of imaging windows on biological processes in longitudinal studies.

