ITSCs cultivated as described above were enzymatically detached using collagenase I and harvested at 300 x g for 10 min. Dissociated cells were transfected with 1 µg pmax GFP vector (Amaxa Biosystems, Switzerland) and Amaxa rat NSC-Nucleofector Kit (Amaxa Biosystems) and the Nucleofector II device (Amaxa Biosystems) according to the manufacturer's information. 5 ml DMEM F-12 (Sigma Aldrich) were added before the sample was centrifuged for 10 min at 300 x g. After discarding the supernatant, cells were cultured as described above. Imaging of transfected cells was performed using a Axio Oberver.D1-Microscope (Carl Zeiss AG, Oberkochen, Germany).
Axio oberver d1 microscope
Manufactured by Zeiss
Sourced in Germany
The Axio Observer.D1 Microscope is an inverted microscope system designed for versatile imaging applications. It features a sturdy stand, a LED illumination system, and a range of optical modules to accommodate various specimen types and techniques. The core function of this microscope is to provide high-quality image acquisition and analysis capabilities.
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2 protocols using axio oberver d1 microscope
1
Transfection of Dissociated Cells
2
Transfection of Dissociated Cells
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ITSCs cultivated as described above were enzymatically detached using collagenase I and harvested at 300 x g for 10 min. Dissociated cells were transfected with 1 µg pmax GFP vector (Amaxa Biosystems, Switzerland) and Amaxa rat NSC-Nucleofector Kit (Amaxa Biosystems) and the Nucleofector II device (Amaxa Biosystems) according to the manufacturer's information. 5 ml DMEM F-12 (Sigma Aldrich) were added before the sample was centrifuged for 10 min at 300 x g. After discarding the supernatant, cells were cultured as described above. Imaging of transfected cells was performed using a Axio Oberver.D1-Microscope (Carl Zeiss AG, Oberkochen, Germany).
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