Journal: Cell systems

Article Title: Geometry dependent arrhythmias in electrically excitable tissues

doi: 10.1016/j.cels.2018.08.013

Figure Lengend Snippet: A) Molecular components of the synthetic excitable tissue. The voltage-gated sodium channel NaV1.5 and inward-rectifying potassium channel Kir2.1 are together sufficient to produce action potentials in response to a depolarizing stimulus. CheRiff is a blue-shifted channelrhodopsin which depolarizes the cells in response to blue light, and BeRST1 is a dye which reports voltage through changes in red fluorescence. Connexin channels (Cxn) introduce electrical coupling between neighboring cells. B) Left: microcontact printing defines patterns of cell growth. The vertical stripes connect in a serpentine pattern outside the field of view. Blue overlay shows regions of optogenetic stimulation. Zero-dimensional “islands” are tested alongside one-dimensional “tracks”. Right: single frame from a movie of BeRST1 fluorescence showing traveling waves in the track region. See Supplementary Movie 1. Scale bars 500 mm. C) Fluorescence recordings of membrane voltage showing that isradipine (10 μM) induces alternans in OS-HEK cells paced at 4 Hz. D) At low pace frequency isradipine OS-HEK (iOS-HEK) cells beat periodically in synchrony with the pacing, but at high pace frequency the cells produce an irregular rhythm.

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, Peptides, and Recombinant Proteins Isradipine Tocris Bioscience 2004 Puromycin ThermoFisher A1113802 Blasticidin ThermoFisher A1113902 Fibronectin Yo Proteins Cat #663 Acrylic acid N-hydroxysuccinimide ester Sigma A8060 Acrylamide solution (40%) Sigma A4058-100ML Acrylamide/Bisacrylamide, (40% solution, 19:1) Sigma A9926-100ML SU-8 3025 Photoresist Microchem SU-8 3025 SU-8 Developer Microchem N/A 3-(Trimethoxysilyl)propyl methacrylate Sigma 2530-85-0 Anhydrous ethanol Sigma 277649-100ML Glacial acetic acid Sigma A6283-100ML Potassium persulfate Sigma 7727-21-1 Experimental Models: Cell Lines Optopatch-Spiking HEK Cells This paper, McNamara et al 2016 32 N/A iCell Cardiomyocytes Cellular Dynamics Kit 01434 Recombinant DNA pENTR-L5-Kir2.1-mCherry-L2 Addgene 32,669 pIRESpuro3-NaV1.5 Johns Hopkins University ChemCORE N/A FCK-Optopatch1 Hochbaum et al 2014 35, Addgene 51695 FCK-CMV-CheRiff-CFP N/A psPAX2 Addgene 12260 VsVg Addgene 12259 Software and Algorithms MATLAB code for image processing and data analysis This paper Supplement MATLAB code for iOS-HEK model simulation and dispersion analysis This paper Supplement MATLAB code for Noble model simulation This paper Supplement Other Firefly Widefield Microscope This paper, Werley et al 2017 39 N/A Silicon Wafers University Wafers 58.52-1 Lithography transparency mask CADArt Services N/A (custom order) Open in a separate window Key Resources Table An engineered excitable cell line becomes arrhythmic when rapidly paced.

Techniques: Fluorescence, Introduce

Journal: Cell systems

Article Title: Geometry dependent arrhythmias in electrically excitable tissues

doi: 10.1016/j.cels.2018.08.013

Figure Lengend Snippet: Key Resources Table

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Chemicals, Peptides, and Recombinant Proteins Isradipine Tocris Bioscience 2004 Puromycin ThermoFisher A1113802 Blasticidin ThermoFisher A1113902 Fibronectin Yo Proteins Cat #663 Acrylic acid N-hydroxysuccinimide ester Sigma A8060 Acrylamide solution (40%) Sigma A4058-100ML Acrylamide/Bisacrylamide, (40% solution, 19:1) Sigma A9926-100ML SU-8 3025 Photoresist Microchem SU-8 3025 SU-8 Developer Microchem N/A 3-(Trimethoxysilyl)propyl methacrylate Sigma 2530-85-0 Anhydrous ethanol Sigma 277649-100ML Glacial acetic acid Sigma A6283-100ML Potassium persulfate Sigma 7727-21-1 Experimental Models: Cell Lines Optopatch-Spiking HEK Cells This paper, McNamara et al 2016 32 N/A iCell Cardiomyocytes Cellular Dynamics Kit 01434 Recombinant DNA pENTR-L5-Kir2.1-mCherry-L2 Addgene 32,669 pIRESpuro3-NaV1.5 Johns Hopkins University ChemCORE N/A FCK-Optopatch1 Hochbaum et al 2014 35, Addgene 51695 FCK-CMV-CheRiff-CFP N/A psPAX2 Addgene 12260 VsVg Addgene 12259 Software and Algorithms MATLAB code for image processing and data analysis This paper Supplement MATLAB code for iOS-HEK model simulation and dispersion analysis This paper Supplement MATLAB code for Noble model simulation This paper Supplement Other Firefly Widefield Microscope This paper, Werley et al 2017 39 N/A Silicon Wafers University Wafers 58.52-1 Lithography transparency mask CADArt Services N/A (custom order) Open in a separate window Key Resources Table An engineered excitable cell line becomes arrhythmic when rapidly paced.

Techniques: Recombinant, Software, Microscopy