3D Transformation Matrix Calculation and Pixel Intensity Normalization for the Dual Focus Tracking System
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We provide details of the dual-focal 3D tracking and pixel intensity calibration (DuFT) system used to record gene expression in targeted areas on the skin of freely moving mice.
The accuracy of the 3D position calculation was determined by placing a scintillator on the calibration disk at various known locations within the recording cage. The height of the disk was varied from the bottom (Z = 0 cm) to the top (Z = 30 cm). The distance from the central axis range from near the center (R = 1 cm) to near to edge (R = 5 cm).
The mean deviation between the known and calculated position was .31 ± 0.16 mm. The maximum deviation was less than .86 mm.
The results indicate that the location of a scintillator within the recording cage imaged with two cameras can be calculated with submillimeter accuracy. We hope that our methods can be applied to improve automatic (even real-time) tracking of various animals in vivo.
KeywordsCircadian rhythm In vivo imaging Fluorescence imaging Multi-camera tracking 3D tracking 3D interpolation
This research was partially supported by Special Expenditures of ‘Reverse Translational Research from Advanced Medical Technology to Advanced Life Science; from Real-time Tracking Technology to Real-time Tracking Life Science’ funded by the Japanese Ministry of Education, Culture, Sports, Science and Technology and was partially supported by a research fund from Research Foundation for Opto-Science and Technology. This work was also supported in part by the Global Station for Medical Science and Engineering, Global Institution for Collaborative Research and Education (GI-CoRE), Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan.
Compliance with Ethical Standards
Conflict of interest
All authors declare that they have no conflict of interest.
Institutional Review Board approval was obtained.
Informed consent was obtained from all individual participants included in the study.
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