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Multiphoton intravital microscopy of the transplanted mouse kidney
G Camirand1, Q Li, A J Demetris
1Surgery Pathology Medicine Immunology, Thomas E. Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA, USA.
Abstract:
Graft outcomes after kidney transplantation continue to be adversely affected by ischemia-reperfusion injury and rejection. High-resolution, real-time imaging of the transplanted kidney could shed valuable insights into these dynamic processes, but such methodology has not been established. Here we describe a technique for intravital imaging of the transplanted mouse kidney using multiphoton fluorescence microscopy. The technique enabled real-time, high-resolution imaging and quantitation of renal filtration, cell death, leukocyte adhesion and capillary blood flow after transplantation. Using this technique, we found that brief graft ischemia associated with the transplantation procedure led to a rapid decline in renal filtration accompanied by a significant increase in microvascular leakage and renal tubular epithelial cell death within the first 3 h after transplantation. No significant changes in leukocyte adhesion or capillary blood flow were observed during the same time period. This report establishes multiphoton fluorescence microscopy as a sensitive tool for simultaneously studying functional and structural perturbations that occur in the mouse kidney after transplantation and for investigating the migration of leukocytes to the graft.
Insights
New multiphoton microscopy allows real-time imaging of transplanted kidneys, revealing rapid filtration decline and cell death post-transplant due to ischemia-reperfusion injury. This technique aids in studying kidney graft function and leukocyte migration.
Area of Science:
- Nephrology
- Transplantation Biology
- Medical Imaging
Background:
- Kidney graft outcomes are negatively impacted by ischemia-reperfusion injury and rejection.
- Established methods for real-time, high-resolution imaging of transplanted kidneys are lacking.
- Understanding dynamic processes in transplanted kidneys is crucial for improving graft survival.
Purpose of the Study:
- To develop and validate a technique for intravital, high-resolution imaging of transplanted mouse kidneys.
- To investigate early functional and structural changes in transplanted kidneys using multiphoton fluorescence microscopy.
- To assess the utility of this technique for studying ischemia-reperfusion injury and immune responses in kidney grafts.
Main Methods:
- Intravital multiphoton fluorescence microscopy of transplanted mouse kidneys.
- Real-time imaging and quantitation of renal filtration, cell death, leukocyte adhesion, and capillary blood flow.
- Analysis of early post-transplantation events within the first 3 hours.
Main Results:
- The developed technique provides real-time, high-resolution imaging capabilities.
- Brief graft ischemia caused a rapid decrease in renal filtration within 3 hours post-transplantation.
- Significant increases in microvascular leakage and renal tubular epithelial cell death were observed.
- No significant changes in leukocyte adhesion or capillary blood flow were detected in the early phase.
Conclusions:
- Multiphoton fluorescence microscopy is a sensitive tool for simultaneous study of functional and structural changes in transplanted kidneys.
- The technique allows for real-time monitoring of early post-transplant events, including ischemia-reperfusion injury.
- This methodology can be used to investigate leukocyte migration and graft response to transplantation.
