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Long Term Intravital Multiphoton Microscopy Imaging of Immune Cells in Healthy and Diseased Liver Using CXCR6.Gfp Reporter Mice
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Imaging liver biology in vivo using conventional confocal microscopy
Pedro E Marques1, Maísa M Antunes1, Bruna A David1
1Laboratório de Imunobiofotônica, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Brasil.
Nature Protocols
|January 9, 2015
Summary
This study details adapting a laser-scanning microscope for live animal imaging, enabling detailed visualization of liver cells in vivo. The protocol offers rapid preparation for extended intravital microscopy (IVM) of cellular function.
Area of Science:
- Cell Biology
- Microscopy Techniques
- Immunology
Background:
- Intravital microscopy (IVM) is crucial for understanding cell biology in live animals.
- The liver's central role in metabolism and immunity necessitates advanced imaging techniques.
Purpose of the Study:
- To adapt a conventional laser-scanning microscope for versatile intravital imaging.
- To establish a protocol for liver confocal IVM in mice to study cellular morphology and function.
- To provide guidelines for simultaneous in vivo staining of multiple cells and molecules.
Main Methods:
- Surgical procedures for liver confocal IVM in mice.
- Adaptation of a low-cost laser-scanning microscope.
- Optimization of antibody and probe doses for in vivo staining.
Main Results:
- Successful implementation of liver confocal IVM allowing visualization of hepatocytes, endothelial cells, and leukocytes.
- Demonstration of analyzing cellular morphology and function in native liver environment.
- Protocol allows up to 4 hours of imaging after 20 minutes of preparation.
Conclusions:
- The developed protocol provides a versatile and cost-effective method for live imaging of liver cells.
- This technique offers valuable insights into liver physiology, pathology, and cell-cell interactions.
- The protocol is adaptable for imaging other abdominal organs like adipose tissue, spleen, and intestines.
