Related Experiment Video
Updated: Oct 3, 2025
Combining Multiplex Fluorescence In Situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections
Published on: June 25, 2021
A new phenotype identification method with the fluorescent expression in cross-sectioned tails in Thy1-GCaMP6s
Shutaro Kobayashi1,2, Kazunori O'Hashi1,3,4, Keisuke Kaneko1,5
1Department of Pharmacology, Nihon University School of Dentistry.
Purpose:
Unless the phenotype of the transgenic mice is distinguishable, genotyping in each mouse is required prior to experiments. This study aimed to establish a new identification method for the phenotype in Thy1-GCaMP6s transgenic mice to reduce the cost and time.
Methods:
Tail biopsies (2 mm) were performed under general anesthesia with isoflurane in 3 to 4-week-old mice. Then, the resected tail was cut again with a sharp razor, and the cross-sections were observed with two-photon microscopy (excitation wavelength = 940 nm). The emitted light was split into green and red light by a dichroic mirror (570 nm) with bandpass filters (495-540 nm for green, 575-645 nm for red).
Results:
Two types of expressed fluorescent pattern were found in the tail tissue: the presence of green fluorescent structures (type 1) and the absence of the structures (type 2). Cortical imaging confirmed that type 1 expressed the cortical GCaMP6s, while type 2 did not.
Conclusion:
These results suggest that observation of the cross-sectioned tail in Thy1-GCaMP6s mice enabled to identify the phenotype within approximately 10 min/mouse, which reduces the cost and time for genotyping.
