Membrane Staining of C2C12 Myoblasts

C2C12 cells (American Type Culture Collection, CRL-1772; RRID: CVCL-0188) were cultured as described previously (12 (link)). For microscopy, myoblasts were plated on glass-bottom dishes (IBIDI, 81158) coated with laminin 111 (Sigma-Aldrich, L2020). Four days after differentiation induction, cells were stained with CellMask Green Plasma Membrane Stain (5 μg/ml; Thermo Fisher Scientific, C37608) for 7 min and imaged live on a Zeiss Cell Observer SD confocal microscope (Carl Zeiss) using a 63× oil objective [numerical aperture (NA) 1.40] and a QImaging ROLERA EM-C2 electron-multiplying charge-coupled device (EMCCD) camera. CellMask Green Plasma Membrane Stain was excited with a 488-nm laser line, and the respective emission was collected with an FE01-520/35 emission filter.

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Kwan H.L., Chan Z.C., Bi X., Kutkowska J., Prószyński T.J., Chan C.B, & Lee C.W. (2023). Nerve-independent formation of membrane infoldings at topologically complex postsynaptic apparatus by caveolin-3. Science Advances, 9(24), eadg0183.

Publication 2023

laminin 111 CellMask Green Plasma Membrane Stain Zeiss Cell Observer SD confocal microscope

Cell Confocal microscope Device Dishes Electron Laminin Microscopy Myoblasts Plasma membrane Stain

Corresponding Organization : Hong Kong Baptist University

Other organizations : Hong Kong University of Science and Technology, Łukasiewicz Research Network – PORT Polish Center for Technology Development

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Variable analysis

independent variables

Differentiation induction

dependent variables

Myoblast morphology (visualized with CellMask Green Plasma Membrane Stain)

control variables

C2C12 cell line (American Type Culture Collection, CRL-1772; RRID: CVCL-0188)
Culture conditions (as described previously in reference 12)
Plating of myoblasts on glass-bottom dishes coated with laminin 111
Staining with CellMask Green Plasma Membrane Stain (5 μg/ml, 7 min)
Imaging on Zeiss Cell Observer SD confocal microscope (63× oil objective, NA 1.40, QImaging ROLERA EM-C2 EMCCD camera)
Excitation with 488-nm laser line, emission collected with FE01-520/35 filter

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