SH-SY5Y cells transfected with GFP-ATG13 using TransIT-2020 reagent (#MIR5400; Mirus) were seeded at a density of 1.0 × 104 cells on μ-Slide 8 Well high (ibidi, Gräfelfing, Germany). After a 24-hr incubation with 2 μM Abe, 100 nM Vac in the growth medium, or HBSS, live-cell imaging was conducted using a 60 × objective lens equipped on a LSM780 confocal laser microscope (Carl Zeiss, Germany). Signal intensity per cell was quantified using ImageJ and the Analyse Particles plugin (a constant threshold for all images per experiment was applied) of Fiji software [14 (link)]. At least 41 cells in each of the three independent experiments were subjected to counting.
μ slide 8 well high
Manufactured by Ibidi
Sourced in Germany
The μ-Slide 8 Well high is a cell culture slide with 8 individual wells designed for microscopic analysis. It provides a standardized and reproducible environment for cell-based assays.
Automatically generated - may contain errors
Lab products found in correlation
3 protocols using μ slide 8 well high
1
Quantifying autophagy in GFP-ATG13 cells
2
Imaging Endosome Dynamics in HeLa Cells
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HeLa cells stably expressing mCherry-2 × FYVE and LAMP1-GFP were seeded at a density of 1.0 × 104 cells on μ-Slide 8 Well high (ibidi). After a 24-hr stimulation with 2 μM Abe or 100 nM Vac, live-cell imaging was conducted using a 60 × objective lens equipped on a LSM780 confocal laser microscope (Carl Zeiss). After setting thresholds for the red (mCherry-2 × FYVE) and green (LAMP1-GFP) signals, the signal intensity of mCherry-2 × FYVE was calculated by the Analyse Particles plugin of Fiji software [14 (link)]. To measure the signal intensity of the double-positive area for mCherry-2 × FYVE and LAMP1-GFP, red and green binary images were overlapped using the paste control “AND”, and signal intensity was calculated by the Analyse Particles plugin. The ratio of mCherry-2 × FYVE co-localized to LAMP1-GFP was calculated from the signal intensity double-positive for red and green divided by the total red signal intensity. At least 23 cells in each of the three independent experiments were subjected to counting.
3
GFP-2xFYVE Imaging in HeLa Cells
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HeLa cells stably expressing GFP-2 × FYVE were seeded at a density of 1.0 × 104 cells on μ-Slide 8 Well high (ibidi). After a 24-hr stimulation with 2 μM Abe or 100 nM Vac with or without 1 μM WM, live-cell imaging was conducted using a 60 × objective lens equipped on a LSM780 confocal laser microscope (Carl Zeiss). Signal intensity per cell was quantified using ImageJ and the Analyse Particles plugin (a constant threshold for all images per experiment was applied) of Fiji software [14 (link)]. At least 20 cells in each of the three independent experiments were subjected to counting.
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