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(A) Experimental tagging techniques: (i) Ventral epithelium - A fluorescent lysotracker dye stains the acidic granules present in the lipophil cells, and provides a dense tagging of the entire ventral epithelium at large fields of view (∼3 mm) (Methods). (ii) Dorsal epithelium - We developed an assay to coat the surface of the epithelium with sticky fluorescent microspheres / microbeads. This provides a coarse-grained tagging (1 bead per 8 cells) of the entire dorsal epithelium at large fields of view (∼6 mm) and enables high-speed (10 fps) and long duration (∼1-5 hours) imaging (Methods). Right Insets display control experiments demonstrating that microspheres bind on cell membrane. (B) Computational data analysis techniques: We employ Flowtrace for visualization of particle trajectories, Particle tracking for quantitative non-affine motion analysis, and Particle Image <t>Velocimetry</t> to measure velocity fields and internal strain rate.
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(A) Experimental tagging techniques: (i) Ventral epithelium - A fluorescent lysotracker dye stains the acidic granules present in the lipophil cells, and provides a dense tagging of the entire ventral epithelium at large fields of view (∼3 mm) (Methods). (ii) Dorsal epithelium - We developed an assay to coat the surface of the epithelium with sticky fluorescent microspheres / microbeads. This provides a coarse-grained tagging (1 bead per 8 cells) of the entire dorsal epithelium at large fields of view (∼6 mm) and enables high-speed (10 fps) and long duration (∼1-5 hours) imaging (Methods). Right Insets display control experiments demonstrating that microspheres bind on cell membrane. (B) Computational data analysis techniques: We employ Flowtrace for visualization of particle trajectories, Particle tracking for quantitative non-affine motion analysis, and Particle Image <t>Velocimetry</t> to measure velocity fields and internal strain rate.
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(A) Experimental tagging techniques: (i) Ventral epithelium - A fluorescent lysotracker dye stains the acidic granules present in the lipophil cells, and provides a dense tagging of the entire ventral epithelium at large fields of view (∼3 mm) (Methods). (ii) Dorsal epithelium - We developed an assay to coat the surface of the epithelium with sticky fluorescent microspheres / microbeads. This provides a coarse-grained tagging (1 bead per 8 cells) of the entire dorsal epithelium at large fields of view (∼6 mm) and enables high-speed (10 fps) and long duration (∼1-5 hours) imaging (Methods). Right Insets display control experiments demonstrating that microspheres bind on cell membrane. (B) Computational data analysis techniques: We employ Flowtrace for visualization of particle trajectories, Particle tracking for quantitative non-affine motion analysis, and Particle Image <t>Velocimetry</t> to measure velocity fields and internal strain rate.
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(A) Experimental tagging techniques: (i) Ventral epithelium - A fluorescent lysotracker dye stains the acidic granules present in the lipophil cells, and provides a dense tagging of the entire ventral epithelium at large fields of view (∼3 mm) (Methods). (ii) Dorsal epithelium - We developed an assay to coat the surface of the epithelium with sticky fluorescent microspheres / microbeads. This provides a coarse-grained tagging (1 bead per 8 cells) of the entire dorsal epithelium at large fields of view (∼6 mm) and enables high-speed (10 fps) and long duration (∼1-5 hours) imaging (Methods). Right Insets display control experiments demonstrating that microspheres bind on cell membrane. (B) Computational data analysis techniques: We employ Flowtrace for visualization of particle trajectories, Particle tracking for quantitative non-affine motion analysis, and Particle Image <t>Velocimetry</t> to measure velocity fields and internal strain rate.
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(A) Experimental tagging techniques: (i) Ventral epithelium - A fluorescent lysotracker dye stains the acidic granules present in the lipophil cells, and provides a dense tagging of the entire ventral epithelium at large fields of view (∼3 mm) (Methods). (ii) Dorsal epithelium - We developed an assay to coat the surface of the epithelium with sticky fluorescent microspheres / microbeads. This provides a coarse-grained tagging (1 bead per 8 cells) of the entire dorsal epithelium at large fields of view (∼6 mm) and enables high-speed (10 fps) and long duration (∼1-5 hours) imaging (Methods). Right Insets display control experiments demonstrating that microspheres bind on cell membrane. (B) Computational data analysis techniques: We employ Flowtrace for visualization of particle trajectories, Particle tracking for quantitative non-affine motion analysis, and Particle Image Velocimetry to measure velocity fields and internal strain rate.

Journal: bioRxiv

Article Title: Motility induced fracture reveals a ductile to brittle crossover in the epithelial tissues of a simple animal

doi: 10.1101/676866

Figure Lengend Snippet: (A) Experimental tagging techniques: (i) Ventral epithelium - A fluorescent lysotracker dye stains the acidic granules present in the lipophil cells, and provides a dense tagging of the entire ventral epithelium at large fields of view (∼3 mm) (Methods). (ii) Dorsal epithelium - We developed an assay to coat the surface of the epithelium with sticky fluorescent microspheres / microbeads. This provides a coarse-grained tagging (1 bead per 8 cells) of the entire dorsal epithelium at large fields of view (∼6 mm) and enables high-speed (10 fps) and long duration (∼1-5 hours) imaging (Methods). Right Insets display control experiments demonstrating that microspheres bind on cell membrane. (B) Computational data analysis techniques: We employ Flowtrace for visualization of particle trajectories, Particle tracking for quantitative non-affine motion analysis, and Particle Image Velocimetry to measure velocity fields and internal strain rate.

Article Snippet: We employed Particle Image Velocimetry (PIV) analysis (PIVlab package in MATLAB) to quantify the flow-fields in the dorsal layer sticky-microbeads time-lapse datasets in large width (13×13mm square) confined PDMS milli-fluidic chips with variable fields-of-view.

Techniques: Imaging, Control, Membrane