Transmission electron microscopy was used to view the vacuole and mitochondria patch (vCLAMP). The strains were cultured in YPD (containing 5 mM H2O2) for 1 h, and then harvested, washed twice by PBS. The cells were suspended in 2.5% glutaraldehyde fixative and fixed at 4 °C for 12 h. Then cells were washed three times by PBS. The cells were then collected by centrifugation, 1% osmic acid was added, and the cells were suspended and fixed at room temperature for 1 h. The cells were collected by centrifugation and washed three times by PBS for 10 min each time. Cells were fixed by gradient ethanol dehydration. The cells were collected by centrifugation and embedded. The embedded samples were cut into 50–100 nm tissue sections using an ultramicrotome. The cuts were then transferred onto a copper mesh and dried. The samples were observed by a transmission electron microscope (Tecnai G2 F-20, FEI Company, Hillsboro, OR, USA).
Immunolabeling was performed using anti-GFP antibody (MBL598, 1:100) for 2 h at room temperature followed by Goat Anti-Rabbit IgG/Gold (10 nm) (Solarbio, 1:100 dilution) for 30 min at room temperature. The samples were further stained with uranium acetate and citric acid, dried, and observed by a transmission electron microscope (FEI Company, Hillsboro, OR, USA).
Immunolabeling was performed using anti-GFP antibody (MBL598, 1:100) for 2 h at room temperature followed by Goat Anti-Rabbit IgG/Gold (10 nm) (Solarbio, 1:100 dilution) for 30 min at room temperature. The samples were further stained with uranium acetate and citric acid, dried, and observed by a transmission electron microscope (FEI Company, Hillsboro, OR, USA).