Cytation 5 plate reader imager

Manufactured by Agilent Technologies

The Cytation 5 is a multi-mode microplate reader and imaging system from Agilent Technologies. It is capable of performing absorbance, fluorescence, and luminescence detection. The Cytation 5 can be used for a variety of applications including cell-based assays, protein quantification, and high-content imaging.

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Lab products found in correlation

Prestoblue reagent, by Thermo Fisher Scientific (2 mentions) Celltiter glo viability assay, by Promega (1 mentions) Matrigel, by Greiner (1 mentions) Cck 8, by Dojindo Laboratories (1 mentions) 384 well plate, by Greiner (1 mentions) Celltiter glo 2.0 viability assay, by Promega (1 mentions) Lsm 510 meta microscope, by Zeiss (1 mentions) Ros glo h2o2, by Promega (1 mentions) Jc 10, by Abcam (1 mentions) Celltiter glo, by Promega (1 mentions)

4 protocols using cytation 5 plate reader imager

Quantitative Viability Assays for hiPSC-CMs

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Day 30–35 post-differentiation hiPSC-CMs were plated on Matrigel at 25,000 cells/well of a 384-well plate (Greiner Bio-One). Cells were treated with TKIs at 0–100 μM for 72 hours unless otherwise specified. Immunostaining qualitatively assessed cell viability per previous protocols (8 (link)). For quantitative viability measurements, cells were treated with CellTiter-Glo Viability Assay (Promega), CCK8 (Dojindo), or PrestoBlue reagent (Life Technologies) per manufacturer-recommended procedures. High-throughput imaging and viability assays were conducted using a Cytation 5 plate reader/imager (BioTek Instruments). Prism (GraphPad) was utilized for curve fitting, LD₅₀ calculations, and statistical analysis.

Sharma A., Burridge P.W., McKeithan W.L., Serrano R., Shukla P., Sayed N., Churko J.M., Kitani T., Wu H., Holmström A., Matsa E., Zhang Y., Kumar A., Fan A.C., del Álamo J.C., Wu S.M., Moslehi J.J., Mercola M, & Wu J.C. (2017). High-Throughput Screening of Tyrosine Kinase Inhibitor Cardiotoxicity with Human Induced Pluripotent Stem Cells. Science translational medicine, 9(377), eaaf2584.

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hiPSC-CM Differentiation Assay

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For 96-well hiPSC-CM differentiation assays, hiPSCs were plated in Matrigel-coated 96-well plates at 1000 cells per well and allowed to adhere for 4 days. The hiPSCs were subsequently treated with bioactive lipids at the indicated concentrations and durations and assessed following day 8 of the chemically-defined hiPSC-CM differentiation protocol. Immunostaining using previously-published protocols was conducted to qualitatively assess cell viability and cardiomyocyte differentiation efficiency^{45 (link)}. Fluorescence intensity and cell number was quantified using ImageJ software. For quantitative viability measurements, cells were treated with CellTiter-Glo 2.0 Viability Assay (Promega) or PrestoBlue reagent (Life Technologies) per manufacturer-recommended procedures. 96-well imaging and viability assays were conducted using a Cytation 5 plate reader/imager (BioTek Instruments). Prism (GraphPad) was utilized for graph generation and statistical analysis. Confocal imaging was performed using a Zeiss LSM 510Meta microscope (Carl Zeiss) using Zen software.

Sharma A., Zhang Y., Buikema J.W., Serpooshan V., Chirikian O., Kosaric N., Churko J.M., Dzilic E., Shieh A., Burridge P.W., Wu J.C, & Wu S.M. (2018). Stage-specific Effects of Bioactive Lipids on Human iPSC Cardiac Differentiation and Cardiomyocyte Proliferation. Scientific Reports, 8, 6618.

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Plate-based Assays for iPSC-CM

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Human iPSC-CMs plated on 96-well plates were subjected to plate-based assays after 5 days of iron ± drug treatment using a Cytation 5 plate reader/imager (BioTek). ROS-Glo H₂O₂ (Promega) was used for ROS detection, JC-10 (Abcam) was used for mitochondrial membrane potential, and Cell Titer Glo (Promega) was used for cell viability assay. All assays were performed according to the manufacturer’s instructions.

Rhee J.W., Yi H., Thomas D., Lam C.K., Belbachir N., Tian L., Qin X., Malisa J., Lau E., Paik D.T., Kim Y., Choi B.S., Sayed N., Sallam K., Liao R, & Wu J.C. (2020). Modeling Secondary Iron Overload Cardiomyopathy with Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes. Cell reports, 32(2), 107886.

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Imaging-based Viability Assay for iPSC-CMs

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iPSC-CMs plated on 96-well plates were treated with iron ± drugs for 5 days. Cells were incubated with 1 μM Calcein-AM and Hoechst 33342 (NucBlue Live ReadyProbes Reagent; 2 drops for 1 ml of media) for 30 min at 37°C. Stained cells were washed three times with PBS and fluorescence images were taken and analyzed by Cytation 5 plate reader/imager (BioTek).

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