Cryo-EM and TEM Imaging of N-AuNSs-Treated Cells

Cells were ready to be examined by cryo-electron microscopy (cryo-EM) and TEM after 24 h of incubation with N-AuNSs. Plunge-freezing of HeLa cells cultured on fibronectin-coated R 2/2 Quantifoil gold EM finder grids (Electron Microscopy Sciences, USA) for cryo-EM was performed as described.^{31,32 (link)} Frozen grids were viewed under the FEI Tecnai G2 Spirit twin (FEI) at 120 kV in cryo-conditions.
Pelleted cells were prepared for TEM as described.^{33,34 (link)} Ultrathin sections were cut using a Leica ultramicrotome (Leica, Austria) and post-stained with uranyl acetate and lead citrate. The grid was viewed under a Zeiss LEO912AB TEM (Carl Zeiss, Germany) at an accelerating voltage of 120 kV.

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Jung D.W., Ro H.J., Kim J., Kim S.I., Yi G.R., Lee G, & Jun S. (2019). Biophysical restriction of growth area using a monodispersed gold sphere nanobarrier prolongs the mitotic phase in HeLa cells. RSC Advances, 9(64), 37497-37506.

Publication 2019

Tecnai G2 Spirit twin Leica ultramicrotome Zeiss LEO912AB TEM

Cells Citrate Cryo electron microscopy Electron microscopy Fibronectin Frozen Gold Hela cells Twin Ultramicrotome Uranyl acetate

Corresponding Organization : Government of the Republic of Korea

Other organizations : Sungkyunkwan University, Suwon Research Institute

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Variable analysis

independent variables

Incubation of HeLa cells with N-AuNSs for 24 h

dependent variables

Examination of HeLa cells by cryo-electron microscopy (cryo-EM)
Examination of HeLa cells by transmission electron microscopy (TEM)

control variables

Plunge-freezing of HeLa cells cultured on fibronectin-coated R 2/2 Quantifoil gold EM finder grids
Viewing of frozen grids under the FEI Tecnai G2 Spirit twin (FEI) at 120 kV in cryo-conditions
Preparation of pelleted cells for TEM as described in the references
Cutting of ultrathin sections using a Leica ultramicrotome and post-staining with uranyl acetate and lead citrate
Viewing of the grid under a Zeiss LEO912AB TEM (Carl Zeiss, Germany) at an accelerating voltage of 120 kV

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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