Aug 27, 2024
Bioluminescence Imaging
- 1Department of Neurological Surgery, Weill Cornell Medical College, New York, NY 10065;
- 2Aligning Science Across Parkinson’s (ASAP) Collaborative Research Network, Chevy Chase, MD 20815, USA
Protocol Citation: Santiago Unda, Michael G. Kaplitt 2024. Bioluminescence Imaging. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov191r2lr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: August 27, 2024
Last Modified: September 23, 2024
Protocol Integer ID: 106521
Keywords: ASAPCRN, imaging, bioluminscence
Funders Acknowledgement:
Aligning Science Across Parkinson's
Grant ID: 020608
Abstract
This protocol describes a bioluminescence stepwise imaging protocol in mice with AAV-luciferase
expression in the vagal afferents using the Ivis Lumina series III imaging system.
Materials
- D-luciferin potassium salt (Xenogen)
- Isoflurane, USP (Covetrus)
Preparation and In vivo imaging
Preparation and In vivo imaging
Induce anesthesia with isoflurane 2-3%.
Shave the neck, thoracic and abdominal area.
Prepare D-luciferin potassium salt (Xenogen) stock prepared in phosphate buffered saline (PBS) at 15mg/mL.
Inject D-luciferin solution (150mg/kg) intraperitoneal.
Image mice in supine position at 5-, 10- and 15-minutes post-luciferin injection.
Software protocol and analysis
Software protocol and analysis
Living imaging software (Ivis imaging) was configured to luminescent imaging mode with an exposure time of 00:00:00 and 5-minute delay.
Post-acquisition, the total flux of photons/radiance per second (s) in each pixel in the ROI area, in a square
centimeter (cm2) of the tissue. This data is then multiplied by one steradian (sr), which result in average radiance displayed as (photons/s/cm2/sr).