Wide-Field Calcium and Flavoprotein Autofluorescence Imaging in Living Mice
- 1Teikyo University
Abstract
Wide-field imaging (WFI) is a mesoscopic approach for monitoring cortex-wide activity with high temporal resolution and a broad field of view. Owing to its simple optical configuration and compatibility with chronic preparations, WFI has become an important tool in systems neuroscience and disease-model research. In this chapter, we describe practical protocols for chronic transcranial WFI in mice using two complementary optical signals: genetically encoded calcium indicators (GCaMP) and endogenous flavoprotein autofluorescence. Calcium imaging provides a robust readout of neuronal population activity, whereas flavoprotein imaging reflects mitochondrial redox dynamics and cellular metabolic demand. We detail procedures for animal preparation, skull clearing, headplate implantation, macroscope assembly, synchronized sensory stimulation, triggered image acquisition, and MATLAB-based data analysis. The analysis workflow includes {Delta}F/F normalization, reference-based signal correction, and artifact reduction, followed by trial averaging, atlas registration, and region-of-interest analysis. Because imaging is performed through the intact skull, the protocol enables repeated longitudinal measurements in the same animal over extended periods. This approach is reproducible, cost-effective, and adaptable to studies of cortical physiology and neurological disorders.
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BibTeX
@misc{yoshida2026widefield,
title = {{Wide-Field Calcium and Flavoprotein Autofluorescence Imaging in Living Mice}},
author = {T. Yoshida},
howpublished = {bioRxiv},
year = {2026},
month = may,
doi = {10.64898/2026.05.14.725112},
url = {https://www.biorxiv.org/content/10.64898/2026.05.14.725112v3},
note = {CC BY-NC-ND},
}
RIS
TY - UNPB
TI - Wide-Field Calcium and Flavoprotein Autofluorescence Imaging in Living Mice
AU - T. Yoshida
PY - 2026
DA - 2026/05/14
PB - bioRxiv
DO - 10.64898/2026.05.14.725112
UR - https://www.biorxiv.org/content/10.64898/2026.05.14.725112v3
AB - Wide-field imaging (WFI) is a mesoscopic approach for monitoring cortex-wide activity with high temporal resolution and a broad field of view. Owing to its simple optical configuration and compatibility with chronic preparations, WFI has become an important tool in systems neuroscience and disease-model research. In this chapter, we describe practical protocols for chronic transcranial WFI in mice using two complementary optical signals: genetically encoded calcium indicators (GCaMP) and endogenous flavoprotein autofluorescence. Calcium imaging provides a robust readout of neuronal population activity, whereas flavoprotein imaging reflects mitochondrial redox dynamics and cellular metabolic demand. We detail procedures for animal preparation, skull clearing, headplate implantation, macroscope assembly, synchronized sensory stimulation, triggered image acquisition, and MATLAB-based data analysis. The analysis workflow includes {Delta}F/F normalization, reference-based signal correction, and artifact reduction, followed by trial averaging, atlas registration, and region-of-interest analysis. Because imaging is performed through the intact skull, the protocol enables repeated longitudinal measurements in the same animal over extended periods. This approach is reproducible, cost-effective, and adaptable to studies of cortical physiology and neurological disorders.
ER -