A total of 150,000 cells were seeded on 25 mm high-precision No. 1.5 borosilicate coverslips (Marienfeld) in 6 well plates (ThermoFisher Scientific) 1 day before transfection in DMEM w/o phenol red (see cell culture). Each chamber was transfected with 2 µg of the plasmid pMD-Vim-Dreiklang17 (link) using Lipofectamine 3000 (ThermoFisher Scientific) according to manufacturer’s instructions and imaged about 24 h post transfection. Before imaging, the coverslip was incubated with 600 nM Mitotracker Deep Red FM in DMEM w/o phenol red for ~30 min at 37 °C and 5% CO2, followed by 10 min incubation of ~10 µg ml−1 WGA-AbberiorFlip565 in Hanks balanced salt solution w/MgCl2 and CaCl2 (HBSS, Gibco). After 1× washing in HBSS the samples were imaged in HBSS at RT for up to 30 min. For imaging, we preswitch Dreiklang using 488 nm light for 10 s before starting data acquisition.
No 1.5 borosilicate coverslips
Manufactured by Marienfeld
No. 1.5 borosilicate coverslips are thin, flat glass sheets commonly used as a support for microscope samples. They are made from borosilicate glass, a type of glass known for its durability and resistance to thermal and chemical stress. These coverslips provide a transparent, flat surface for mounting and protecting specimens during microscopic examination.
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Lab products found in correlation
Lipofectamine 3000, by Thermo Fisher Scientific (2 mentions)
6 well plate, by Thermo Fisher Scientific (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
Lab tek 2 chambered cover slides, by Thermo Fisher Scientific (1 mentions)
12 well plate, by Thermo Fisher Scientific (1 mentions)
2 protocols using no 1.5 borosilicate coverslips
1
Visualizing Vimentin Dynamics in Cells
2
Immunofluorescence Staining of Tubulin in COS-7 Cells
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Fixed COS-7 cells were prepared as described in 19 (link) . Primary anti-tubulin antibody (clone B-5-1-2 ascites fluid, Sigma-Aldrich) was used at 1:100-1:200 dilution, secondary donkey anti-mouse-AbberiorFlip565
(preparation see below) was used at 1:100-1:200 dilution.
COS-7 cells were cultured at 37 °C and 5 % CO2 using DMEM high glucose w/o phenol red (4.5 g l -1 glucose) supplemented with 4 mM L-gluthamine, 10 % fetal bovine serum and 1× penicillin-streptomycin (all gibco ® , Thermo Fisher Scientific). Cells were seeded in Lab-tek ® II chambered cover slides (nunc) or on 18 mm high-precision No. 1.5 borosilicate coverslips (Marienfeld) in 12 well plates (Thermo Fisher Scientific)
1-2 days before fixation in DMEM high glucose w/o phenol red (see above).
The fixation and labelling protocol is similar as described by Chazeau et al. 24 (link)
(preparation see below) was used at 1:100-1:200 dilution.
COS-7 cells were cultured at 37 °C and 5 % CO2 using DMEM high glucose w/o phenol red (4.5 g l -1 glucose) supplemented with 4 mM L-gluthamine, 10 % fetal bovine serum and 1× penicillin-streptomycin (all gibco ® , Thermo Fisher Scientific). Cells were seeded in Lab-tek ® II chambered cover slides (nunc) or on 18 mm high-precision No. 1.5 borosilicate coverslips (Marienfeld) in 12 well plates (Thermo Fisher Scientific)
1-2 days before fixation in DMEM high glucose w/o phenol red (see above).
The fixation and labelling protocol is similar as described by Chazeau et al. 24 (link)
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