Synergy neo hts multimode microplate reader

Manufactured by Agilent Technologies

Sourced in United States

The Synergy neo HTS multimode microplate reader is a compact and versatile instrument designed for high-throughput screening and multi-mode detection. It is capable of measuring absorbance, fluorescence, and luminescence in a variety of microplate formats.

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Lab products found in correlation

Ros assay kit, by Beyotime (3 mentions) Cytotox 96 non radioactive cytotoxicity assay kit, by Promega (2 mentions) Fluorescein isothiocyanate (fitc), by Merck Group (2 mentions) Alexa fluor 555, by Thermo Fisher Scientific (2 mentions) Mmp assay kit, by Beyotime (2 mentions) Renilla luciferase assay lysis buffer, by Promega (2 mentions) Renilla luciferase assay system, by Promega (2 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Rneasy mini kit, by Qiagen (1 mentions) 96 well plate, by Thermo Fisher Scientific (1 mentions) Cck 8 cell counting kit, by Vazyme (1 mentions) Lactic dehydrogenase, by Merck Group (1 mentions) Pyruvate kinase from rabbit muscle, by Merck Group (1 mentions) Myokinase from chicken muscle, by Merck Group (1 mentions) Dcfh da ros assay kit, by Beyotime (1 mentions) Nano glo live cell assay system, by Promega (1 mentions) Multidrop combi reagent dispenser, by Thermo Fisher Scientific (1 mentions) Echo 550, by Beckman Coulter (1 mentions) Prism 7, by GraphPad (1 mentions) Taqman universal pcr master mix, by Thermo Fisher Scientific (1 mentions)

42 protocols using synergy neo hts multimode microplate reader

Alveolus-Chip Surfactant and Cytokine Assay

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(which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this preprint this version posted August 4, 2020. ; https://doi.org/10.1101/2020.08.02.233338 doi: bioRxiv preprint PBS (200µl) was washed through the epithelial surface of the Open-Top Alveolus-Chip by pipetting, and collected, for measurement of Surfactant C levels using an ELISA kit for Human Surfactant Protein C (LifeSpanBioScience: LS-F12438), following the manufacturer's protocol. The color reaction was measured at 450nm with a Synergy NEO HTS Multi-Mode microplate reader (BioTek). Samples were stored at -80°C until analysis. Similarly, the presence of cytokines (IL-6, IL-8 and MPC-1) was detected from medium samples by Human TLR-induced Cytokines II: Microbial-induced Multi-Analyte ELISArray Kit (Qiogen: MEH-008A) following the manufacturer's protocol. The color reaction was measured at 450nm with a Synergy NEO HTS Multi-Mode microplate reader (BioTek). Samples were stored at -80°C until analysis.

Varone A., Nguyen J.K., Leng L., Barrile R., Sliz J., Lucchesi C., Wen N., Gravanis A., Hamilton G.A., Karalis K, & Hinojosa C.D. (2021). A novel organ-chip system emulates three-dimensional architecture of the human epithelia and the mechanical forces acting on it. Biomaterials, 275.

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Evaluating Cell Viability via MTT Assay

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Cell viability was evaluated by MTT assay as described by us previously 11 (link). LNCaP and DU145 cells were cultured in 96 well plates in triplicates overnight at 37°C. The cells were incubated with different concentrations of PSN-A for 24 h with or without 10 µM BAPTA-AM. Following treatments, cells were incubated with 10 µL MTT reagent (5mg/mL) in the dark at 37°C for 4 h. Finally, medium was carefully removed and 150µL DMSO was added into each well to dissolve farmazan crystals. The plates were shaked gently to mix the color. The absorbance was then measured at 570 nm by a microplate reader (Synergy neo HTS multimode microplate reader, BioTek). The percentage of cell viability was calculated using following formula.

He Y., Khan M., Yang J., Yao M., Yu S, & Gao H. (2018). Proscillaridin A induces apoptosis, inhibits STAT3 activation and augments doxorubicin toxicity in prostate cancer cells. International Journal of Medical Sciences, 15(8), 832-839.

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MTT Cell Viability Assay

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The parental and STAT3 knockdown A549 cells were cultured in a 96-well plate for 24 h. The cells were incubated with 10 μL MTT reagent (5 mg/mL) at 37°C for 4 h. Subsequently, 150 μL DMSO was added to dissolve formazan crystals. The absorbance was measured at 570 nm by a microplate reader (Synergy neo HTS multimode microplate reader, BioTek).

Maryam A., Mehmood T., Yan Q., Li Y., Khan M, & Ma T. (2018). Proscillaridin A Promotes Oxidative Stress and ER Stress, Inhibits STAT3 Activation, and Induces Apoptosis in A549 Lung Adenocarcinoma Cells. Oxidative Medicine and Cellular Longevity, 2018, 3853409.

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Cell Viability Assay Protocol

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Cell viability was examined by measurement of released lactate dehydrogenase (LDH) from microfluidic device medium outflow, spent medium from cells cultured on tissue culture plates, cell lysate (positive control), or fresh cell culture medium (negative control) by using the CytoTox 96^® NonRadioactive Cytotoxicity Assay Kit (Promega) per manufacturer’s protocol. Absorbance was measured with a Synergy NEO HTS Multi-Mode microplate reader (BioTek).

Musah S., Mammoto A., Ferrante T.C., Jeanty S.S., Hirano-Kobayashi M., Mammoto T., Roberts K., Chung S., Novak R., Ingram M., Fatanat-Didar T., Koshy S., Weaver J.C., Church G.M, & Ingber D.E. (2017). Mature induced-pluripotent-stem-cell-derived human podocytes reconstitute kidney glomerular-capillary-wall function on a chip. Nature biomedical engineering, 1, 0069.

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Cell Viability Assay Protocol

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Microfluidic Evaluation of Renal Filtration

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Urinary clearance of inulin and albumin was evaluated by using cell-lined microfluidic devices subjected to both fluid flow and cyclic strain, and cultured for a minimum of 8 days. For filtration studies, both the top and bottom channels of the device were perfused with Complete Medium with CultureBoost-R. Cell culture medium supplemented with a mixture of 10 μg/mL inulin conjugated to FITC (Sigma-Aldrich) and 100 μg/mL albumin conjugated to Alexa Fluor 555 (ThermoFisher Scientific) was continuously perfused through the microvascular channel for 6 h while applying cyclic strain. The fluorescence intensity of the outflow from each channel was measured by using a Synergy NEO HTS Multi-Mode microplate reader (BioTek). The amount of inulin or albumin filtered from the microvascular to the urinary channel was calculated by using the equation for renal clearance:

Urinary clearance = ([U] \times UV) / [P]

Where [U] = urinary concentration, UV = urinary volume, and [P] = plasma or microvascular concentration. The percent urinary clearance was calculated from a ratio of urinary clearance to UV.

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Quantifying Colony Formation in HepG2 Cells

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HepG2 cells exposed to different concentrations of STM for 4 h were seeded and cultured in DMEM with 1% FBS in 6-well culture plate (300/well). After two weeks on appearance of colonies, cells were washed twice with PBS, fixed with 4% paraformaldehyde (PFA) for 20 minutes. Cells were then stained with crystal violet for 15 minutes at room temperature. Extra stain was removed by washing with PBS until background was clear. Stained colonies were rinsed by methanol to dissolve crystal violet and O.D. was calculated at 595 nm by fluorescent spectrophotometer (Synergy neo HTS multi-mode microplate reader, BioTek).

Mehmood T., Maryam A., Tian X., Khan M, & Ma T. (2017). Santamarine Inhibits NF-кB and STAT3 Activation and Induces Apoptosis in HepG2 Liver Cancer Cells via Oxidative Stress.. Journal of Cancer, 8(18), 3707-3717.

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CCK-8 Assay for Cell Viability

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Cell viability was measured by CCK-8 assay kit according to the manufacturer’s instructions. Briefly, MCF-7 and MDA-MB-231 cells were seeded in 96-well microtiter plates. After overnight incubation at 37°C, cells were treated with different indicated concentrations of Brv-A in presence or absence of NAC (5 mM) for 24 h. Following treatments, 10 µL of CCK-8 solution was added into each well and kept for 3 h at 37°C. Optical density was recorded at wavelength of 450 nm by using fluorescence microplate reader (Synergy neo HTS multimode microplate reader, BioTek). All experiments were performed in triplicate and cell viability was calculated using following formula.
Cell viability (%) = (A450 sample – A450 blank)/(A450 control – A450 blank) × 100
where “sample” shows optical density (OD) of drug treated cells with CCK-8 solution while “blank” represents OD value of culture medium with CCK-8 but without cells.

Saleem M.Z., Nisar M.A., Alshwmi M., Din S.R., Gamallat Y., Khan M, & Ma T. (2020). Brevilin A Inhibits STAT3 Signaling and Induces ROS-Dependent Apoptosis, Mitochondrial Stress and Endoplasmic Reticulum Stress in MCF-7 Breast Cancer Cells. OncoTargets and therapy, 13, 435-450.

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Intracellular GSH Quantification

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The intracellular GSH was measured using a glutathione reduced assay kit (Nanjing, Jiancheng, China). The cells were cultured in six-well plates in triplicates and treated with 10 and 20 µM Brevilin A for 4 hours. The cells were collected, washed with PBS, and samples were prepared in accordance with the manufacturer’s instructions. Finally, the intracellular GSH was measured using a fluorescent spectrophotometer (Synergy Neo HTS Multi-Mode Microplate Reader, BioTek) at a wavelength of 405 nm. The GSH contents were expressed as nmol GSH/mg protein.

Wang J., Li M., Cui X., Lv D., Jin L., Khan M, & Ma T. (2018). Brevilin A promotes oxidative stress and induces mitochondrial apoptosis in U87 glioblastoma cells. OncoTargets and therapy, 11, 7031-7040.

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Cell Viability Assay of PSD-A in Breast Cancer

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PSD-A was dissolved in DMSO with its final concentration of 0.5% and DMSO treated cells with the same concentration were used as control. A CCK-8 kit was used to determine cell viability according to the manufacturer’s instructions. Briefly, MCF-7 and MDA-MB-231 were seeded in 96-well microtiter plates with density of 3 ×10³ cells/well/100 µL and treated with different concentrations of PSD-A for 24h as indicated. Following treatments, 10 µL of CCK-8 solution was added to each well and kept for 3h at 37°C and OD was recorded at a wavelength of 450 nm by using a fluorescence microplate reader (Synergy neo HTS multimode microplate reader, BioTek). All experiments were performed in triplicate and cell viability was calculated using following formula.
where “sample” shows optical density (OD) of drug treated cells with CCK-8 solution while “blank” represents OD value of culture medium with CCK-8 but without cells.

Saleem M.Z., Alshwmi M., Zhang H., Din S.R., Nisar M.A., Khan M., Alam S., Alam G., Jin L, & Ma T. (2020). Inhibition of JNK-Mediated Autophagy Promotes Proscillaridin A- Induced Apoptosis via ROS Generation, Intracellular Ca+2 Oscillation and Inhibiting STAT3 Signaling in Breast Cancer Cells. Frontiers in Pharmacology, 11, 01055.

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