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Bioluminescence measurements in mice using a skin window
Qin Huang1, Victor Acha, Raylon Yow
1University of Texas Health Science Center, Department of Physiology and Sam and Ann Barshop Institute for Longevity and Aging Studies, San Antonio, Texas 78245, USA.
Journal of Biomedical Optics
|November 13, 2007
Summary
A novel plastic window in mice allows accurate measurement of internal bioluminescence, overcoming tissue absorption issues. This technique enables precise monitoring of bioluminescent cells and biological processes within living animals.
Area of Science:
- Biomedical Engineering
- Molecular Biology
- Animal Models
Background:
- Accurate measurement of bioluminescence in living animals is crucial for biosensor applications.
- Tissue absorption of light, particularly below 600 nm, impedes reliable measurements.
- Existing methods struggle with quantifying light emission from internal organs.
Purpose of the Study:
- To develop a novel method for measuring bioluminescence from internal organs in vivo.
- To overcome the limitations of tissue light absorption in animal models.
- To enable precise quantification of bioluminescent signals for biosensing applications.
Main Methods:
- A plastic window was surgically implanted in the skin/body wall of mice.
- Bioluminescent cells coexpressing firefly luciferase (FLuc), Renilla luciferase (RLuc), and GFP2 fusion proteins were transplanted into the kidney.
- Bioluminescence and bioluminescence resonance energy transfer (BRET) were measured using fiber optics, a photon counter, and specific optical filters after substrate administration.
- Vasopressin was administered to assess the system's response.
Main Results:
- The plastic window facilitated accurate measurement of bioluminescence from internal kidney cells.
- Distinct light emissions from FLuc and RLuc were distinguishable using optical filters.
- Bioluminescence resonance energy transfer (BRET) was successfully detected and quantified.
- Intraperitoneal injection of vasopressin resulted in a significant increase in BRET.
Conclusions:
- The combination of a skin/body wall window and fiber optic measurement provides an advantageous method for rapid and accurate light measurement from internal organs.
- This technique overcomes tissue absorption limitations, enabling robust in vivo bioluminescence studies.
- The developed system is suitable for cell-based biosensor applications and monitoring dynamic biological processes in real-time.
