Short report
"Color Timer" mice: visualization of neuronal differentiation with fluorescent proteins
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* Corresponding author: Hideyuki Okano hidokano@sc.itc.keio.ac.jp
1 Department of Physiology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo 160-8582, Japan
2 Laboratory for Cell Function Dynamics, Advanced Technology Development Core, Brain Science Institute (BSI), Institute of Physical and Chemical Research (RIKEN), 2-1 Hirosawa, Wako-shi, Saitama 351-0198, Japan
3 Current address: Departamento de Histologia e Embriologia, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro (UFRJ), Rio de Janeiro, Brazil
Molecular Brain 2010, 3:5 doi:10.1186/1756-6606-3-5
Published: 2 February 2010Additional files
Additional file 1:
Spectral characteristics of KOr and EGFP. The numerical data for the KOr spectra were obtained at https://ruo.mbl.co.jp/product/flprotein/ko.html/ webcite.
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Transition from orange to green fluorescence in the cerebral cortex of embryonic nestin/KOr (line #70) - DCX-EGFP double Tg mice. Cerebral cortices of E14.5 double Tg mice with line #70 of nestin/KOr were isolated, manually cut with a knife, embedded in a collagen (Nitta Gelatin) matrix, and time-lapse imaged with an LSM5 PASCAL fluorescence microscope (Zeiss). The 543-nm HeNe laser and 488-nm Argon laser were used to excite KOr and EGFP, respectively. The cell indicated by the arrow is a representative example of the orange-to-green transition.
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Transition from orange to green fluorescence in nestin/KOr (line #55) - DCX-EGFP double Tg mice. The cerebral cortex of an E14.5 double Tg mouse embryo with line #55 of nestin/KOr was time-lapse imaged.
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Transition from orange to green fluorescence in neurospheres derived from cerebral cortex of embryonic nestin/KOr (Line #70) - DCX-EGFP double Tg. Cerebral cortices of E14.5 double Tg mouse embryos with line # 70 of nestin/KOr were isolated, dissociated into individual cells by mechanical pipetting, cultured for several days in a medium containing EGF and bFGF, and time-lapse imaged.
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