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Synergy htx multi mode microplate reader

Manufactured by Agilent Technologies
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The Synergy HTX Multi-Mode Microplate Reader is a versatile laboratory instrument designed for various detection modes. It can perform absorbance, fluorescence, and luminescence measurements on microplates.

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Lab products found in correlation

Take3 micro volume plate, by Agilent Technologies (8 mentions) Prestoblue cell viability reagent, by Thermo Fisher Scientific (6 mentions) Dual luciferase reporter assay system, by Promega (6 mentions) Invitrap spin universal rna mini kit, by Stratec (5 mentions) Prism 8, by GraphPad (5 mentions) Tri reagent, by Merck Group (4 mentions) High capacity cdna reverse transcription kit, by Thermo Fisher Scientific (4 mentions) Dual luciferase reporter assay kit, by Promega (4 mentions) Bright glo luciferase assay system, by Promega (4 mentions) Celltiter glo luminescent cell viability assay reagent, by Promega (4 mentions) Biostack3 microplate stacker, by Agilent Technologies (3 mentions) Staurosporine, by Merck Group (3 mentions) Bovine serum albumin (bsa), by Fujifilm (3 mentions) P nitrophenyl phosphate pnpp, by Merck Group (3 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (3 mentions) Bradford assay, by AppliChem (3 mentions) Prestoblue, by Thermo Fisher Scientific (3 mentions) Alamarblue reagent, by Thermo Fisher Scientific (3 mentions) Fitc dextran, by Merck Group (3 mentions) X tremegene 9 dna transfection reagent, by Roche (3 mentions)

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Microplate reader (6925 citations) Synergy HTX (873 citations) Synergy HTX Multi-Mode Reader (387 citations) Synergy microplate reader (229 citations) Synergy HTX microplate reader (95 citations) Synergy HTX Multi-Reader (24 citations) Synergy HTX reader (19 citations) Synergy HTX Multi-Mode (12 citations) HTX Microplate Reader (11 citations) HTX multi-mode reader (4 citations)

398 protocols using synergy htx multi mode microplate reader

1

Osteoblastic Differentiation and Mineralization

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Cell differentiation into the osteoblastic lineage was evaluated by measuring the alkaline phosphatase (ALP) activity as a differentiation marker. For this purpose, the same extracts obtained in the cell proliferation assay were used for quantifying the ALP activity with a SensoLyte pNPP Alkaline Phosphatase Assay Kit (AnaSpec Inc., Fremont, CA, USA). Enzymatic activity was evaluated spectrophotometrically at 405 nm for each specified time point using a Synergy HTX multi-mode microplate reader (Bio-Tek Instruments, USA), and the results were normalized versus their corresponding cell number obtained in the proliferation assay. Mineralization was assessed after culturing the cells for 28 days on scaffolds. Then, the cells were fixed in 4% paraformaldehyde solution in PBS for 15 min and rinsed with distilled water. Calcium deposits were stained with 500 µL/scaffold of 40 mM Alizarin Red S (Sigma-Aldrich, USA) for 20 min by gentle shaking. Excess dye was removed by several washings with distilled water, and the staining was extracted by incubation in 10% cetylpyridinium chloride with 10 mM NaH2PO4. Supernatants were measured spectrophotometrically at 570 nm using a Synergy HTX multi-mode microplate reader (Bio-Tek Instruments, USA), and the results were normalized versus their corresponding cell number obtained in the proliferation assay.
Guillem-Marti J., Vidal E., Girotti A., Heras-Parets A., Torres D., Arias F.J., Ginebra M.P., Rodriguez-Cabello J.C, & Manero J.M. (2023). Functionalization of 3D-Printed Titanium Scaffolds with Elastin-like Recombinamers to Improve Cell Colonization and Osteoinduction. Pharmaceutics, 15(3), 872.
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2

Quantitative Analysis of Cell Viability

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For each extract, the rate of HAC/GFP loss was accurately measured independently using Synergy™ HTX Multi-Mode Microplate Reader (BioTek, United States) and BD Accuri C6 (BD Biosciences, United States) for a double check. For Synergy™ HTX Multi-Mode Microplate Reader (BioTek,United States), 1 × 105 cells were seeded on a 96-well plate and depicted 24 h after extract treatment under 4х magnification in 5 × 5 fields of view. The number of GFP-positive and GFP-negative cells were counted in each field with threshold 7,000. The cell size was restricted from 5 μm up to 100 μm. For flowcytometry experiment using BD Accuri C6 (BD Biosciences, United States), 4 × 105 cells per well were seeded on a 24-well plate with 500 μl of the culture medium and 10 μg/ml of Blasticidin S. Then cells were harvested by trypsin-treatment and resuspended in PBS containing 3 µM of DRUQ7. All the samples were vortexed immediately prior to flow cytometry examination using BD Accuri C6 (BD Biosciences, United States), the Fluorescence of GFP-positive cells was measured by the 488 nm laser and detected at 510 nm. The death cells were counted by DRUQ7 fluorescence excited by the 640 nm laser and detected at 722 nm. Samples were acquired in at least three separate triplicates for 30 s or 1.5 × 104 events (at minimum).
Goncharov N.V., Kovalskaia V.A., Romanishin A.O., Shved N.A., Belousov A.S., Tiasto V.S., Gulaia V.S., Neergheen V.S., Rummun N., Liskovykh M., Larionov V., Kouprina N, & Kumeiko V.V. (2022). Novel assay to measure chromosome instability identifies Punica granatum extract that elevates CIN and has a potential for tumor- suppressing therapies. Frontiers in Bioengineering and Biotechnology, 10, 989932.
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3

Mucosal Disaccharidase Activity Measurement

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The disaccharidase activity of mucosal scrapings was assessed using a modified glucose oxidase peroxidase enzyme system. Jejunal mucosal tissue samples were homogenized in saline and centrifuged (2,500 × g, 4 °C, 10 min) to produce the supernatant. The enzyme sucrase, lactase and maltase activities were analyzed using commercial kits (Jiancheng Bioengineering Institute, Nanjing, China) and Synergy HTX Multi-Mode Microplate Reader (BioTek, Vermont, USA) following the manufacturer's instructions. The concentration of protein was also analyzed also using commercial bicinchoninic acid assay kits (Jiancheng Bioengineering Institute, Nanjing, China) and Synergy HTX Multi-Mode Microplate Reader (BioTek, Vermont, USA) according to the manufacturer's instructions. Throughout this paper, disaccharidase activity has been expressed as units per milligram protein of the mucosal sample.
Wang L., Tan X., Wang H., Wang Q., Huang P., Li Y., Li J., Huang J., Yang H, & Yin Y. (2020). Effects of varying dietary folic acid during weaning stress of piglets. Animal Nutrition, 7(1), 101-110.
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4

Biomarker Profiling in Plasma

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The contents of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP), albumin (ALB), globulin (GLB), glucose, triglyceride (TG), total cholesterol (TCH), blood urea nitrogen (BUN), insulin-like growth factor 1 (IGF1), growth hormone (GH), malondialdehyde (MDA), IgG, IgA, and IgM and the activities of the T-AOC, SOD, GSH-px, and CATU in the plasma were measured by spectrophotometric methods (Jiancheng Biological Engineering Research Institute, Nanjing, China) using a Synergy HTX Multi-Mode Microplate Reader (BioTek Instruments Inc., Winooski, Vt., USA). The IL-1β, IL-2, IL-4, and TNF-α contents were measured by ELISA kits (Cloud-Clone Corporation, Houston, USA) using a Synergy HTX Multi-Mode Microplate Reader (BioTek Instruments Inc., Winooski, Vt., USA).
Wu S., Li X., Chen X., Zhu Y, & Yao J. (2021). Optimizing the growth and immune system of dairy calves by subdividing the pre-weaning period and providing different milk volumes for each stage. Animal Nutrition, 7(4), 1296-1302.
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5

Glycan Binding Preference of Avian and Human Influenza Viruses

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The binding preference of A/SW1/18 for avian- and human-specific virus receptors was confirmed using a solid-phase direct binding assay, as described previously (Koo et al., 2018 (link)). In brief, 10 μg of fetuin (Sigma-Aldrich, MO, USA) was coated and incubated overnight at 4°C and blocked with 5% bovine serum albumin (BSA) in PBS at room temperature for 1 h. After blocking, the plates were washed four times with PBS and incubated with 64 hemagglutinating units (HAU) of each virus at 4°C overnight. After virus removal, the plate was washed as described above and incubated with 0.1 ml of each biotinylated glycan per well at different concentrations at 4°C for 3 h. Glycan binding was detected by adding horseradish peroxidase-conjugated streptavidin (Invitrogen, Carlsbad, CA, USA) followed by 3,3,5,5-tetramethylbenzidine substrate (Sigma-Aldrich), and the optical density was measured at 450 nm using a Synergy HTX multi-mode microplate reader (BioTek, VT, USA). A low-pathogenic H5N2 avian influenza virus, A/Aquatic Bird/Korea/CN2/2009 (A/CN2/09), with a binding preference for the avian receptor, was used as a control (Koo et al., 2018 (link)).
Zanin M., Le T.B., Na W., Kang J.A., Kwon H.J., Hwang J., Ga E.H., Wong S.S., Cho H.J., Song D., Kim H.K., Jeong D.G, & Yoon S.W. (2023). Potential for transmission of naturally mutated H10N1 avian influenza virus to mammalian hosts and causing severe pulmonary disease. Frontiers in Microbiology, 14, 1256090.
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6

Cell Viability Assay with Tetrazolium MTS

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Prior to each experiment, cells were counted and diluted to the respective density of 3.0 × 105 cells/mL. The viability of cells was assessed by means of tetrazolium MTS assay using the CellTiter96® Aqueous One Solution Cell Proliferation Assay (Promega GmbH, Walldorf, Germany) according to the manufacturer´s instructions. The cells were treated with appropriate amounts of compounds of interest or corresponding vehicle (vehicle control: DMSO (<0.01%) was the vehicle control for all FA additions, and water was the vehicle control for all 5-Me-THF additions). Assays were performed in triplicate in 96-well plates. Three hours before the indicated time points, 10 µL of MTS reagent was added to each well containing 100 µL of cell suspension. Absorbance at 492 nm was measured after 3 h incubation on an automated microplate reader, the Synergy™ HTX Multi-Mode Microplate Reader (BioTek Instruments, Inc., Winooski, VT, USA). The relative cell viability of treated cells was calculated by subtracting the negative control, which contained no cells, and normalized to vehicle-treated control cells.
Vidmar Golja M., Šmid A., Karas Kuželički N., Trontelj J., Geršak K, & Mlinarič-Raščan I. (2020). Folate Insufficiency Due to MTHFR Deficiency Is Bypassed by 5-Methyltetrahydrofolate. Journal of Clinical Medicine, 9(9), 2836.
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7

MTT Assay for Cell Viability

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Cells were seeded at 2 × 105 cells/ml in different wells of four 96-well plate and incubated for 4 days. Each day, a plate was removed and processed. A total of 50 µl of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) substrate was added and incubated for 1 h. Cell supernatant was removed carefully and replaced with 100 µl of DMSO. Absorbance reading at 570 nm was measured using a Synergy HTX Multi-Mode microplate reader (BioTek Instruments, Winooski, VT).
Cheok Y.Y., Tan G.M., Fernandez K.C., Chan Y.T., Lee C.Y., Cheong H.C., Looi C.Y., Vadivelu J., Abdullah S, & Wong W.F. (2021). Podoplanin Drives Motility of Active Macrophage via Regulating Filamin C During Helicobacter pylori Infection. Frontiers in Immunology, 12, 702156.
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8

Quantifying Fungal Transcription Mutant Fitness

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In total, 2 × 104 spores/mL from each of the 484 members of the transcription knockout library were added to 1× RPMI 1640 medium, 165 mM MOPS buffer (pH 7.0), 2% glucose in each well of a CytoOne 96-well plate with 0.002 mg/L olorofim (n = 4). Plates were incubated at 37°C for 48 h. Fitness was calculated by dividing the optical density of respective null mutants by the MFIG001 control. Relative fitness in olorofim was calculated by dividing fitness in olorofim with general growth fitness of the transcription factor null mutants using the same microculture conditions in 1× RPMI 1640 medium, 165 mM MOPS buffer (pH 7.0), and 2% glucose without olorofim (n = 4). Optical density was measured at 600 nm on a Synergy HTX Multi-Mode Microplate Reader (BioTek, Winooski, VT).
van Rhijn N., Hemmings S., Storer I.S., Valero C., Bin Shuraym H., Goldman G.H., Gsaller F., Amich J, & Bromley M.J. (2022). Antagonism of the Azoles to Olorofim and Cross-Resistance Are Governed by Linked Transcriptional Networks in Aspergillus fumigatus. mBio, 13(6), e02215-22.
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9

Lactate Dehydrogenase Cytotoxicity Assay

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EpiDerm™ cell media were collected from flavor chemical and ECEAR treatments and immediately assayed using the OPS Diagnostics Lactate Dehydrogenase Protocol. Tris-HCl (Sigma-Aldrich, T-3253) and Tris-base (Sigma-Aldrich, T4661) were combined to make TRIS, pH 8. Iodonitrotetrazolium chloride (INT, Sigma I-8377) dissolved in DMSO (Sigma D-8779), phenazine methosulfate (PMS, Sigma P-9625), nicotinamide adenine dinucleotide (NAD, Sigma N-0632), and lithium lactate (Sigma L-1500) were prepared before use and stored at −20°C. 50 μL of 100mM TRIS were added to 50 μl of 50 mM lithium lactate and 50 μl of PMS, INT, NAD. 50 μl of sample were added to each well and after 5 minutes, the absorbance was read on a Synergy™ HTX Multi-Mode Microplate Reader (BioTek, VT). The cytotoxicity was calculated:
Abs(x)−Abs(NegativeCN)Abs(PositiveCN)−Abs(NegativeCN)x100
Khachatoorian C., Luo W., McWhirter K.J., Pankow J.F, & Talbot P. (2021). E-Cigarette Fluids and Aerosol Residues Cause Oxidative Stress and an Inflammatory Response in Human Keratinocytes and 3D Skin Models. Toxicology in vitro : an international journal published in association with BIBRA, 77, 105234.
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10

Hydrogen Peroxide Quantification in Cell Cultures

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The Promega ROS-Glo™ H2O2 Assay (Madison, WI) was used to measure hydrogen peroxide in culture media, culture media plus refill fluid, and culture media plus refill fluid and cells. According to Promega, this kit is specific for hydrogen peroxide. The non-lytic protocol provided by the manufacturer was followed. Control reactions without cells were used to determine spontaneous hydrogen peroxide generation. The groups studied were medium only, medium with positive control (10 μM menadione), medium with 1% Dewberry Cream, and medium with 1% Churrios. Cells were seeded on a 96 well plate at 1000 cells/well and allowed to attach for 24 hrs., after which they were incubated in medium only, medium with the positive control, medium with 1% Dewberry Cream, and medium with 1% Churrios for 24 hrs. 20μl of H2O2 Substrate solution were added to all samples at 18 hrs. (final 6 hrs. of treatment). When treatment was done, 50μl of medium from each sample well was combined with 50μl of ROS-Glo™ Detection Solution in a separate plate and incubated for 20 minutes at room temperature. A Synergy™ HTX Multi-Mode Microplate Reader (BioTek, VT) was used to read relative luminescence units (RLU) (gain = 135, bottom optics). The experiment was done in triplicate, and each sample was done in duplicate; average luminescence is reported.
Khachatoorian C., Luo W., McWhirter K.J., Pankow J.F, & Talbot P. (2021). E-Cigarette Fluids and Aerosol Residues Cause Oxidative Stress and an Inflammatory Response in Human Keratinocytes and 3D Skin Models. Toxicology in vitro : an international journal published in association with BIBRA, 77, 105234.
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