Multicolor Fluorescence Microscopy Dynamics

mCherry and EGFP fluorescence were acquired in a temperature and CO₂-controlled SP8 Leica confocal microscope with a 63× multi-immersion objective. 60 Z-stacks of six images each were acquired (0.34 μm Z-distance, 15 s intervals, 15 min). Average intensity projections were assembled into QuickTime movies (10 fps, 150x speed). F-actin dynamics in DAOY cells stably expressing LA-EGFP [19 (link)] were recorded with a temperature- and CO₂ controlled Zeiss Axio Observer epifluorescence microscope using a glycerol 63x objective (30 s intervals, 60 min).

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Tripolitsioti D., Kumar K.S., Neve A., Migliavacca J., Capdeville C., Rushing E.J., Ma M., Kijima N., Sharma A., Pruschy M., McComb S., Taylor M.D., Grotzer M.A, & Baumgartner M. (2018). MAP4K4 controlled integrin β1 activation and c-Met endocytosis are associated with invasive behavior of medulloblastoma cells. Oncotarget, 9(33), 23220-23236.

Publication 2018

SP8 Leica confocal microscope Axio Observer epifluorescence microscope

Acquired temperature Cells Confocal microscope F actin Fluorescence Glycerol Immersion Microscope

Corresponding Organization : University Children's Hospital Zurich

Other organizations : University Hospital of Zurich, University of Toronto, Hospital for Sick Children, SickKids Foundation

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Variable analysis

independent variables

Temperature
CO2 concentration

dependent variables

MCherry fluorescence
EGFP fluorescence
F-actin dynamics

control variables

Temperature
CO2 concentration
Microscope setup (SP8 Leica confocal, Zeiss Axio Observer epifluorescence)
Objective lens (63× multi-immersion, glycerol 63×)
Cell line (DAOY cells stably expressing LA-EGFP)

controls

Positive control: Not specified
Negative control: Not specified

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