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Pherastar fs plate reader

Manufactured by BMG Labtech
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Sourced in Germany, United Kingdom, United States, France

The PHERAstar FS is a high-performance, multi-mode microplate reader designed for a wide range of applications. It offers rapid and sensitive detection of fluorescence, luminescence, and absorbance signals in 6- to 384-well microplates.

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Lab products found in correlation

Mars data analysis v 3.01r2, by BMG Labtech (10 mentions) Tb streptavidin, by Thermo Fisher Scientific (4 mentions) Bradford assay, by Bio-Rad (4 mentions) Bright glo luciferase assay reagent, by Promega (4 mentions) Passive lysis buffer (plb), by Promega (4 mentions) Nluc substrate furimazine, by Promega (4 mentions) Alamarblue cell viability reagent, by Thermo Fisher Scientific (3 mentions) Celltiter glo 3d cell viability assay, by Promega (3 mentions) 50ss pins, by V&P Scientific (3 mentions) Prism 8, by GraphPad (3 mentions) Celltiter glo reagent, by Promega (2 mentions) Daf fm diacetate, by Thermo Fisher Scientific (2 mentions) Prism 5, by GraphPad (2 mentions) H2dcfda, by Thermo Fisher Scientific (2 mentions) Mars data analysis software, by BMG Labtech (2 mentions) Echo555 acoustic transfer system, by Beckman Coulter (2 mentions) Bravo automated liquid handling, by Agilent Technologies (2 mentions) El406 microplate washer dispenser, by Agilent Technologies (2 mentions) H2dcfda, by Merck Group (2 mentions) Ctg assay, by Promega (2 mentions)

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PHERAstar FS (237 citations) PHERAstar (145 citations) PHERAstar plate reader (142 citations) PHERAstar FS reader (8 citations) Pherastar FS multimode plate reader (5 citations) BMG Pherastar FS plate reader (2 citations)

125 protocols using pherastar fs plate reader

1

Assessing Neuronal Viability and Metabolic Activity

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It is crucial to investigate the effect of any drug on the viability and metabolic activity of the target cells. We assessed the viability of VM neurons and ATP release in response to pregabalin treatment after 3 days of culture. To assess the viability of the VM neurons in the control and pregabalin-treated cultures, we used alamarBlue™ Cell Viability Reagent (Thermo Fisher, USA). We performed the experiment according to the manufacturer’s instructions. After preparing the well plates, the fluorescence was measured using a PHERAstar FS plate reader (BMG LabTech, Ortenberg, Germany).
ATP release was assessed using the CellTiter-Glo® 3D Cell Viability Assay (Promega, Madison, Wisconsin, WI, USA) as an indicator of the metabolic activity status of the cells. Briefly, CellTiter-Glo® Reagent was added in a volume equal to that of the cell culture medium and mixed thoroughly by pipetting up and down 10 times to break down the 3D construct comprising the cells and hydrogel. Plates were then incubated at room temperature for 25 min, and the luminescent signal was read using a PHERAstar FS plate reader (BMG LabTech, Germany).
Alsanie W.F., Alhomrani M., Gaber A., Habeeballah H., Alkhatabi H.A., Felimban R.I., Abdelrahman S., Hauser C.A., Chaudhary A.G., Alamri A.S., Raafat B.M., Alamri A., Anwar S., Alswat K.A., Althobaiti Y.S, & Asiri Y.A. (2022). The Effects of Prenatal Exposure to Pregabalin on the Development of Ventral Midbrain Dopaminergic Neurons. Cells, 11(5), 852.
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2

Evaluating Drug Effects on Neuronal Viability

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It is crucial to assess how a drug affects the metabolic activity and viability of target cells. After three days of culture, we exposed VM neurons to QEPF to measure their survival and ATP release. We determined the viability of VM neurons in untreated and QEPF-treated cells using alamarBlue™ Cell Viability Reagent (ThermoFisher Scientific, Waltham, MA, USA) as per the manufacturer’s instructions. A PHERAstar FS plate reader (BMG LabTech, Ortenberg, Germany) was used to measure fluorescence after the well plates were prepared. ATP release was measured as a marker of the metabolic activity of cells using the CellTiter-Glo® 3D cell viability assay (Promega, Madison, WI, USA). The 3D construct comprised of the cells and hydrogel was thoroughly mixed by pipetting up and down ten times after CellTiter-Glo® Reagent was introduced in an amount similar to the cell culture medium. A PHERAstar FS plate reader (BMG LabTech, Ortenberg, Germany) was used to scan the plates after 25 min of incubation at room temperature to detect the existence of a strong signal. Three wells from each experiment (seven biological replicates for viability and three for ATP release) were analyzed.
Alsanie W.F., Abdelrahman S., Alhomrani M., Gaber A., Alosimi E.A., Habeeballah H., Alkhatabi H.A., Felimban R.I., Hauser C.A., Tayeb H.H., Alamri A.S., Alamri A., Raafat B.M., Alswat K.A., Althobaiti Y.S, & Asiri Y.A. (2022). The Influence of Prenatal Exposure to Quetiapine Fumarate on the Development of Dopaminergic Neurons in the Ventral Midbrain of Mouse Embryos. International Journal of Molecular Sciences, 23(20), 12352.
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3

Screening Maybridge Ro3 1000 Fragment Library

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The Maybridge Ro3 1000 fragment library (Maybridge) screen was performed in black, 384-well plates (PerkinElemer). Displacement of GlcNAcstatin BF was measured by adding 25 μL of 7 nM CpOGA and 1 nM GlcNAcstatin BF in assay buffer (0.1 M Tris-HCl pH 7.4, 150 mM NaCl, 1% DMSO) to assay plates containing 50 nL of a 0.1 M fragment solutions in DMSO, resulting in a final assay concentration of 200 μM. The plates were allowed to stand for 10 minutes in the dark before reading polarization on a Pherastar FS plate reader (BMG Labtech) at excitation and emission wavelengths of 485 nm and 520 nm, respectively. Readings were corrected for background polarization from reactions containing only 1 nM GlcNAcstatin BF in assay buffer and normalised to readings containing 1% DMSO. Fragments displacing GlcNAcstatin BF by ≥ 40% were classified as hits and were advanced for Ki value determination. Competition binding experiments were conducted under the same assay conditions in 0.1 M Tris-HCl pH 7.4, 150 mM NaCl, 2% DMSO and varying concentrations of fragments. Ki values were calculated as described above.
Borodkin V.S., Rafie K., Selvan N., Aristotelous T., Navratilova I., Ferenbach A.T, & van Aalten D.M. (2018). O-GlcNAcase fragment discovery with fluorescence polarimetry. ACS chemical biology, 13(5), 1353-1360.
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4

Bioluminescence Resonance Energy Transfer Assays

Cited 1 time
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The saturation and
competition binding
assays were performed according to the methodology of Stoddart.18 (link) Briefly, the assays were performed on stably
transfected NLucP2Y2-1321N1 cells that had been seeded
24 h prior to the experiment in white Thermo Scientific Matrix 96-well
microplates. The medium in each well was removed and replaced with
HBSS containing apyrase (1 U/mL) and the required concentration of
the fluorescent ligand with or without the competing ligand. Upon
the addition of the fluorescent ligand, the cells were incubated for
1 h at 37 °C without CO2. The NLuc substrate, furimazine
(Promega), was then added to a final concentration of 10 μM,
and the plate was incubated for a further 5 min at 37 °C without
CO2. The luminescence and resulting BRET were measured
using a PHERAstar FS plate reader (BMG Labtech) at room temperature.
For the assays involving 97, sequential measurements
of the filtered light emissions were made at 460 nm (80 nm bandpass)
and >610 nm (long-pass), and the raw BRET ratios were calculated
by
dividing the >610 nm emissions by the 460 nm emissions. For the
assays
involving 98, the measurements were made at 475 nm (30
nm bandpass) and 535 nm (30 nm bandpass), and the raw BRET ratios
were calculated by dividing the 535 nm emissions by the 475 nm emissions.
Conroy S., Kindon N.D., Glenn J., Stoddart L.A., Lewis R.J., Hill S.J., Kellam B, & Stocks M.J. (2018). Synthesis and Evaluation of the First Fluorescent Antagonists of the Human P2Y2 Receptor Based on AR-C118925. Journal of Medicinal Chemistry, 61(7), 3089-3113.
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5

AlamarBlue Assay for Cell Metabolism

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Cell metabolic activity was measured with the AlamarBlue assay (BioRad, Watford, UK) according to the manufacturer’s instructions. Samples were incubated with 10% AlamarBlue at 37°C, 5% CO2 until a colour change was visible. Fluorescence intensity was obtained at 540 nm excitation and 590 nm emission (PHERASTAR FS plate reader; BMG Labtech).
Liu P., Roberts S., Shoemaker J.T., Vukasinovic J., Tomlinson D.C, & Speirs V. (2023). Validation of a 3D perfused cell culture platform as a tool for humanised preclinical drug testing in breast cancer using established cell lines and patient-derived tissues. PLOS ONE, 18(3), e0283044.
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6

In Vitro Protein Expression and TR-FRET Binding Assay

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GST (pcDNA3.1-FLAG-GST) or GST-PXR LBD [pcDNA3.1-FLAG-GST-PXR LBD (WT, W299A, and W299D)] were expressed using the TNT Quick Coupled Transcription/Translation System (Promega); 1 μg of plasmid DNA was used for each 50 μL reaction, and the reactions were incubated at 30°C for 90 min. The TR-FRET binding assay has been described previously [30 (link)], and the protocol was modified for the in vitro protein expression method. Reactions (20 μL) contained 50 mM Tris (pH 7.5), 50 mM NaCl, 0.1 mg/mL bovine serum albumin, 5 nM LanthaScreen Tb-anti-GST Antibody (Thermo Fisher Scientific), 2 μL of protein expression reaction product, 1% DMSO, and the indicated concentrations of BODIPY FL vindoline. Reactions were incubated at RT in black 384-well low-volume assay plates for 30 min, and a PHERAstar FS plate reader (BMG Labtech) was used to detect the TR-FRET signals with the following instrumentation settings: a 340 nm excitation filter, a 100 μs delay time, and a 200 μs integration time. The TR-FRET ratio was expressed as 10,000 × 520 nm/490 nm, and values were normalized by subtracting the GST control signal at each point from the corresponding GST-PXR LBD signals.
Huber A.D., Wright W.C., Lin W., Majumder K., Low J.A., Wu J., Buchman C.D., Pintel D.J, & Chen T. (2020). Mutation of a single amino acid of pregnane X receptor switches an antagonist to agonist by altering AF-2 helix positioning. Cellular and molecular life sciences : CMLS, 78(1), 317-335.
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7

Fluorescence-based Viral RNA Release Assay

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A dried nickel-chelating lipid film was rehydrated in a buffer containing 50 mM HEPES (pH 7.2), 50 mM NaCl, YoPro-1 (ThermoFisher) at a 1:500 ratio (vol/vol), and 1% glucose. RDLs with a lipid concentration of 1 mg/mL were prepared. RNase A (Sigma) was added to achieve a final concentration of 200 µg/mL to eliminate the background signals outside the liposomes. Then, 10 µL of 1 mg/mL virus was incubated with 90 µL of RDLs. The RNA release assay measured the fluorescence signal from YoPro-1 binding with FCV RNA releasing into RDLs. To assess the pH impact on RNA release, the pH was adjusted using 1 M sodium acetate (pH 3.0) and verified with pH paper. Fluorescence measurements were taken every 5 min for 1 h at 37°C using a PHERAstar FS plate reader (BMG Labtech) with excitation and emission wavelengths of 490 nm and 520 nm, respectively. For visualizing the RNA release, the RDL and virus were incubated at different pH conditions for 30 min at 37°C and imaged using an inverted fluorescence microscope (EVOS FL, Life, USA) with a 20 × objective.
Sun W., Wang M., Shi Z., Wang P., Wang J., Du B., Wang S., Sun Z., Liu Z., Wei L., Yang D., He X, & Wang J. (2024). VP2 mediates the release of the feline calicivirus RNA genome by puncturing the endosome membrane of infected cells. Journal of Virology, 98(5), e00350-24.
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8

Measuring cAMP Modulation in Cell Lines

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cAMP experiments
were performed using Flp-In T-REx293 cells induced to express the
receptor of interest or CHO-K1 cells stably expressing the orthologue
of interest. Experiments were carried out using a homogeneous time-resolved
FRET-based detection kit (CisBio, Codolet, France) according to the
manufacturer’s protocol. For the assay cells were plated at
5000 cells/well in low-volume 384-well plates. The ability of agonists
to inhibit 1 μM forskolin-induced cAMP production was assessed
following a preincubation for 15 min with antagonist compounds, then
a further 30 min incubation with agonist compounds. Reactions were
stopped according to the manufacturer’s instructions and the
output was measured with a PHERAstar FS plate reader (BMG Labtech,
Aylesbury, UK).
Jenkins L., Marsango S., Mancini S., Mahmud Z.A., Morrison A., McElroy S.P., Bennett K.A., Barnes M., Tobin A.B., Tikhonova I.G, & Milligan G. (2021). Discovery and Characterization of Novel Antagonists of the Proinflammatory Orphan Receptor GPR84. ACS Pharmacology & Translational Science, 4(5), 1598-1613.
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9

Methamphetamine Effects on EVMN Viability

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We evaluated the viability of EVMNs and ATP release in response to methamphetamine treatment after three days of culture. The alamarBlueTM Cell Viability Assay Reagent kit (Thermo Fisher Scientific) was used to determine the viability of the EVMNs in the control and methamphetamine-treated cultures.
The CellTiter-Glo® 3D Cell Viability Assay (Promega, Madison, Wisconsin, WI, USA) was used to measure ATP release to assess the metabolic activity of the cells following the manufacturer’s instructions. Briefly, CellTiter-Glo® Reagent (Promega) was added in a volume equal to that of the cell culture media in the plate and mixed by pipetting up and down 10 times to break the 3D construct comprising cells and hydrogel. Afterward, the plates were incubated for 25 min at room temperature, and the luminescent signal was recorded using a PHERAstar FS plate reader (BMG LabTech, Ortenberg, Germany).
Alsanie W.F., Abdelrahman S., Felimban R.I., Alkhatabi H.A., Gaber A., Alosimi E.A., Alhomrani M., Habeeballah H., Hauser C.A., S. Alamri A., Althobaiti A., Alsharif A., Alzahrani A.S., Al-Ghamdi M.S., Raafat B.M., Alswat K.A., Althobaiti Y.S, & Asiri Y.A. (2023). The Influence of Prenatal Exposure to Methamphetamine on the Development of Dopaminergic Neurons in the Ventral Midbrain. International Journal of Molecular Sciences, 24(6), 5668.
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10

SRB Assay for Cell Growth Evaluation

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The effect of the different compounds on cell growth was evaluated using the sulforhodamine B (SRB) assay, as previously described [47 (link)]. Briefly, 3000–5000 cells/well were seeded in 96-well plates. After 24 h, cells were incubated with indicated concentrations of each compound for 72 h. Then, the medium was removed and cells were fixed with a trichloroacetic acid solution (10% final concentration) and stained with 0.4% SRB solution in 1% acetic acid for 30 min. Cells were washed three times with 1% acetic acid and SRB was dissolved in 10 mmol/L Tris-HCl solution by gentle shaking. Absorbance at 560 nm was then measured using a PHERAstar FS plate reader (BMG Labtech, Champigny s/Marne, France). Percent growth was calculated as compared to untreated cells and plotted as a function of concentrations. Results are the mean ± sd of three independent experiments.
Corvaglia V., Ait Mohamed Amar I., Garambois V., Letast S., Garcin A., Gongora C., Del Rio M., Denevault-Sabourin C., Joubert N., Huc I, & Pourquier P. (2021). Internalization of Foldamer-Based DNA Mimics through a Site-Specific Antibody Conjugate to Target HER2-Positive Cancer Cells. Pharmaceuticals, 14(7), 624.
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