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Microplate reader

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The Microplate reader is a laboratory instrument used to measure the absorbance, fluorescence, or luminescence of samples in a microplate format. It facilitates rapid and accurate quantification of various biological and chemical assays.

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Lab products found in correlation

Methyl thiazolyl tetrazolium (mtt), by Merck Group (68 mentions) Cell counting kit 8 (cck8), by Dojindo Laboratories (34 mentions) Cck 8, by Dojindo Laboratories (31 mentions) Dimethyl sulfoxide (dmso), by Merck Group (21 mentions) Cck 8 solution, by Dojindo Laboratories (20 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (20 mentions) Cck 8, by Tecan (20 mentions) Cell counting kit 8 cck 8, by Dojindo Laboratories (20 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (20 mentions) Mtt solution, by Merck Group (19 mentions) Cck 8 kit, by Dojindo Laboratories (17 mentions) 96 well plate, by Corning (15 mentions) Cck 8 reagent, by Dojindo Laboratories (14 mentions) Bovine serum albumin (bsa), by Merck Group (12 mentions) Cck 8 assay, by Dojindo Laboratories (12 mentions) Cell counting kit 8 cck 8, by Beyotime (11 mentions) Cck 8, by Beyotime (11 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (11 mentions) Mtt reagent, by Merck Group (9 mentions) Methyl thiazolyl tetrazolium (mtt), by Genview (9 mentions)

Close spelling variants of this product

Safire2 microplate reader (97 citations) Ultra Multifunctional Microplate Reader (45 citations) M200 PRO microplate reader (24 citations) Automatic microplate reader (16 citations) Automated microplate reader (15 citations) Infinite F50 absorbance microplate reader (12 citations) Infinite® F50/Robotic absorbance microplate readers (5 citations) Infinite M200 fluorescence microplate reader (5 citations) Multiskan GO microplate reader (4 citations) In nite M200 microplate reader (4 citations)

2 786 protocols using microplate reader

1

Tissue Calcium, Protein, and Hydroxyproline Analysis

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The tissue calcium content was determined in kidney, heart, aortic, and carotid artery tissue. Total calcium was mobilized by 10% formic acid overnight, followed by a colorimetrical quantification by o-cresolphthalein (Randox Laboratories, Crumlin UK). Absorbance of the supernatant was read at 550 nm on a Tecan microplate reader (Tecan, Männedorf, Switzerland) [29 (link)].
Total protein was assessed by the bicinchoninic acid method (Pierce, Thermo Fisher, Carlsbad, CA, USA) [30 (link)]. Absorbance was measured after 30 min of incubation at 37 °C at 562 nm on a microplate reader (Tecan, Männedorf, Switzerland) [30 (link)].
Hydroxyproline was analyzed after homogenization of cardiac tissue on a ball mill (Qiagen, Germany) and hydrolyzation overnight at 37 °C in 10 N NaOH, followed by neutralization by 10 N HCl. A colorimetric quantification by the reaction of oxidized hydroxyproline with 4-(dimethylamino) benzaldehyde (Sigma Aldrich, St. Louis, MO, USA) was recorded on a microplate reader (Tecan, Männedorf, Switzerland) at 560 nm [31 (link)].
Barros X., Friesen X., Mathias Brandenburg V., Liehn E.A., Steppan S., Kiessling F., Kramann R., Floege J., Krüger T, & Kaesler N. (2023). Magnesium Improves Cardiac Function in Experimental Uremia by Altering Cardiac Elastin Protein Content. Nutrients, 15(6), 1303.
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2

Evaluating HTB Derivatives in BV-2 Cells

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The 2.5 × 104 cells/mL BV-2 cells were seeded in 96-well plate. Cells were pretreated with indicated synthetic HTB derivatives for 1 h, with or without LPS (100 ng/mL) for 24 h. The effect of HMA on BV-2 cell viability was determined, as described previously [34 (link)]. After treatment for the desired time, supernatants were suctioned or removed by flipping the plate. The formazan crystals formed in viable cells were dissolved in DMSO. Optical density was measured at 550 nm using a microplate reader (Tecan Trading AG, Basel, Switzerland) and the values were compared with untreated cells.
The inhibitory effect of synthetic HTB derivatives and HMA (0.1, 1, and 10 µM) on nitrite concentration was determined, as described previously [44 (link)]. Briefly, a standard curve was generated using a range of dilutions of known concentration of sodium nitrite. Approximately 540 nm wavelength was used to measure the absorbance in a microplate reader (Tecan Trading AG).
Song S.Y., Kim I.S., Koppula S., Park J.Y., Kim B.W., Yoon S.H, & Choi D.K. (2020). 2-Hydroxy-4-Methylbenzoic Anhydride Inhibits Neuroinflammation in Cellular and Experimental Animal Models of Parkinson’s Disease. International Journal of Molecular Sciences, 21(21), 8195.
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3

Evaluating DM Inhibition of LPS-Induced Inflammation

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BV-2 cells were seeded at a density of 5 × 104 cells/well and were pretreated with various concentrations of DM (100, 250, and 500 μg/ml) for 1 h, followed by LPS (200 ng/ml) induction for 24 h. 20 μl of MTT (2.0 mg/ml) was added to each well, and after 2 h of incubation at 37°C in 5% CO2, the supernatants were removed from each well, and the formed formazan crystals in viable cells were dissolved in DMSO. The absorbance was determined at 540 nm using a microplate reader (Tecan Trading AG, Basel, Switzerland). The inhibitory effect of DM on NO production was determined as previously described [25 (link)]. BV-2 microglial cells (5 × 104 cells/well) were incubated with LPS (200 ng/ml) in the presence or absence of DM (100, 250, and 500 μg/ml) for 24 h. After 24 h, 100 μl of supernatants was initially collected and assayed for NO release using commercially available Griess reagent (1 vol. 0.1% naphthylethylenediamine and 1 vol. 1% sulfanilamide in 5% H3PO4). Absorbance was determined at 540 nm using a microplate reader (Tecan Trading AG).
Park S.Y., Karthivashan G., Ko H.M., Cho D.Y., Kim J., Cho D.J., Ganesan P., Su-Kim I, & Choi D.K. (2018). Aqueous Extract of Dendropanax morbiferus Leaves Effectively Alleviated Neuroinflammation and Behavioral Impediments in MPTP-Induced Parkinson's Mouse Model. Oxidative Medicine and Cellular Longevity, 2018, 3175214.
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4

Measurement of Mitochondrial and Cellular ROS

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Mitochondrial ROS production was assessed with Mito SOX red probe (Fisher Scientific, New Hampshire, USA) according to the manufactory’s protocol. The cells were seeded in a black 96-well plate. After exposure, the cells were washed with HBSS (Gibco, Fisher Scientific, New Hampshire, USA), and incubated with 5 μM Mito SOX red probe for 15 min. After washing with HBSS, the fluorescence intensity was measured at an excitation wavelength of 510 nm/emission at 590 nm by micro-plate reader (TECAN, Zurich, Switzerland).
Total cellular ROS measurement was evaluated by H2DCFDA according to the manufactory’s protocol (Fisher Scientific, New Hampshire, USA). After washing with PBS, the cells were incubated with 10 μM of DCFDA at 37 °C in dark for 30 min. After washing with HBSS, the fluorescence intensity (excitation = 485 nm; emission = 530 nm) was measured using a micro-plate reader (TECAN, Zurich, Switzerland).
Wu R., Högberg J., Adner M., Ramos-Ramírez P., Stenius U, & Zheng H. (2020). Crystalline silica particles cause rapid NLRP3-dependent mitochondrial depolarization and DNA damage in airway epithelial cells. Particle and Fibre Toxicology, 17, 39.
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5

ReBiL2.0 Assay for RAS Activation

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ReBiL2.0 assay was performed as previously descried 16 . ReBiL cells (U2OS-134-764np or U2OS-134-794p with overexpression of KRAS4b, HRAS or NRAS) were seeded in i) 96-well plates at density of 2x10 4 , and ii) 6-well plates at density of 1x10 6 and allowed to adhere overnight in regular growth media (DMEM/F12, 10% FBS, and 10 µg/mL ciprofloxacin). The next day, cells were then cultured in serum limited media (DMEM/F12, 1% FBS, and 10 µg/mL ciprofloxacin) containing 100 ng/mL doxycycline for 24 hours. Upon termination of the ReBiL assay, i) to measure raw luciferase activity, 300 µM D-luciferin was added to 96-well plate culture and incubate in 37 o C for 30mins and raw luminescent data collected by a Tecan microplate reader; ii) to measure viable cell numbers, CCK-8 assay were performed in the same 96-well plate culture and raw cell number data collected by a Tecan microplate reader; iii) to quantify the 1/2luc fusion proteins, ReBiL cells from 6-well plate culture were harvested with RIPA lysis buffer for protein extraction and western blot was performed for HA-tag and HSP90 expression. Then the ReBiL2.0 score was calculated via the formula:
Tang R., Shuldiner E.G., Kelly M.R., Murray C.W., Hebert J.D., Andrejka L., Tsai M.K., Hughes N.W., Parker M.I., Cai H., Li Y., Wahl G.M., Dunbrack R.L., Jackson P.K., Petrov D.A., & Winslow M.M. (2021). Multiplexed identification of RAS paralog imbalance as a driver of lung cancer growth.
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6

ReBiL2.0 Assay for RAS Activation

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ReBiL2.0 assay was performed as previously descried 16 . ReBiL cells (U2OS-134-764np or U2OS-134-794p with overexpression of KRAS4b, HRAS or NRAS) were seeded in i) 96-well plates at density of 2x10 4 , and ii) 6-well plates at density of 1x10 6 and allowed to adhere overnight in regular growth media (DMEM/F12, 10% FBS, and 10 µg/mL ciprofloxacin). The next day, cells were then cultured in serum limited media (DMEM/F12, 1% FBS, and 10 µg/mL ciprofloxacin) containing 100 ng/mL doxycycline for 24 hours. Upon termination of the ReBiL assay, i) to measure raw luciferase activity, 300 µM D-luciferin was added to 96-well plate culture and incubate in 37 o C for 30mins and raw luminescent data collected by a Tecan microplate reader; ii) to measure viable cell numbers, CCK-8 assay were performed in the same 96-well plate culture and raw cell number data collected by a Tecan microplate reader; iii) to quantify the 1/2luc fusion proteins, ReBiL cells from 6-well plate culture were harvested with RIPA lysis buffer for protein extraction and western blot was performed for HA-tag and HSP90 expression. Then the ReBiL2.0 score was calculated via the formula:
Tang R., Shuldiner E.G., Kelly M.R., Murray C.W., Hebert J.D., Andrejka L., Tsai M.K., Hughes N.W., Parker M.I., Cai H., Li Y., Wahl G.M., Dunbrack R.L., Jackson P.K., Petrov D.A., & Winslow M.M. (2021). Multiplexed identification of RAS paralog imbalance as a driver of lung cancer growth.
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7

Quantifying Acetylcholine and Esterase in Brain Tissue

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The concentration of acetylcholine was determined using Acetylcholine ELISA kit (MybioSource, San Diego, CA, USA). The standard solution and brain tissue homogenates were dispensed into a prepared 96-well plate, and 10 μL of balance solution and 50 μL of conjugate were dispensed into each well, followed by incubation at 37 °C for 1 h. The solution was washed and 100 μL of substrate solution was added to induce color development. The absorbance was measured using a microplate reader (TECAN, Port Melbourne, VIC, Austria) at 450 nm.
The concentration of acetylcholine esterase was determined using an acetylcholine esterase ELISA kit (Cusabio, Balitmore, MD, USA). After all the treatments specified for the assay, TMB substrate solution was added to develop the color and the absorbance was measured at 450 nm using a microplate reader (TECAN, Port Melbourne, VIC, Austria). The activity of acetylcholine esterase in brain tissue homogenates was determined using the acetylcholineesterase assay kit (BioAssay System, Hayward, CA, USA). The enzyme activity was measured at 412 nm at intervals of 2 and 10 min.
Kim H.L., Kim W.K, & Ha A.W. (2019). Effects of Phytochemicals on Blood Pressure and Neuroprotection Mediated Via Brain Renin-Angiotensin System. Nutrients, 11(11), 2761.
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8

Plasma-Induced Oxidative Stress Quantification

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Fully supplemented RPMI1640 medium was loaded with 1 µM singlet oxygen sensor, or PBS was loaded with 1 µM of either the fluorescent redox indicators APF or HPF (life technologies). Both APF and HPF can be oxidized by hydroxyl radicals and peroxynitrite but not hydrogen peroxide whereas only APF is sensitive towards hypochlorous acid29 (link). Two-hundred and fifty microliter was added to each well of a 24-well plate, and wells either received plasma treatment or were left untreated. Subsequently, the liquid was aliquoted into 96-well plates, and fluorescence was acquired using a microplate reader with λex 485 nm and λem 535 nm (Tecan). In order to quantify hydrogen peroxide (H2O2), plasma-treated cell culture medium or plasma-treated chloride-free phosphate buffer was incubated with amplex ultra red (life technologies) according to the vendor’s instructions, and subsequently quantified against an H2O2 standard using a Tecan microplate reader with λex 530 nm and λem 590 nm. Alternatively, cell culture medium was spiked with a known concentration of H2O2 and the scavenging activity of plasma-introduced reactive components on H2O2 was assessed in a similar manner.
Bekeschus S., Wende K., Hefny M.M., Rödder K., Jablonowski H., Schmidt A., Woedtke T.V., Weltmann K.D, & Benedikt J. (2017). Oxygen atoms are critical in rendering THP-1 leukaemia cells susceptible to cold physical plasma-induced apoptosis. Scientific Reports, 7, 2791.
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9

Oxidative Stress Markers Assays

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H2O2 concentrations were determined using a H2O2 assay kit (BioVision, Inc., CA, USA). OxiRed probe reacts with H2O2 to produce a product with color (570 nm) in the presence of horseradish peroxidase. The reaction product was determined by measuring the absorbance at 570 nm using a Sunrise Rainbow RC-R microplate reader (Tecan Austria GmbH, Salzburg, Austria).
NO was assayed using an NO (NO2/NO3) assay kit (NO2/NO3 colorimetric assay kit-C II, Dojindo Laboratories, Kumamoto, Japan). The crude homogenate described previously was heat-treated to remove proteins before use. NO content was calculated based on the amount of NO2/NO3 by measuring the absorbance at 540 nm using a microplate reader (Tecan).
Malondialdehyde (MDA) was determined using a TBARS assay kit (ZeptoMetrix Crop., NY, USA). MDA-thiobarbituric acid adducts formed from the reaction of MDA in the sample were determined by measuring the absorbance at 540 nm using a microplate reader (Tecan).
The carbonyl proteins of control and B12-deficient worms were determined using a modified 2,4-dinitrophenylhydrazine (DNPH) method [20] . Carbonyl content was determined by measuring the absorbance at 380 nm and calculated using a molar absorption coefficient of 21,000 (mol/L)−1 cm−1.
All oxidative stress markers were assayed according to the respective manufacturer's instructions or the cited reference.
Bito T., Misaki T., Yabuta Y., Ishikawa T., Kawano T, & Watanabe F. (2016). Vitamin B12 deficiency results in severe oxidative stress, leading to memory retention impairment in Caenorhabditis elegans. Redox Biology, 11, 21-29.
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10

Monolayer and 3D Viability Assays

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For monolayer culture viability assays, CNO-treated cells were plated on Matrigel-coated 96-well plates at a density of 50,000 cells/well for 48 h prior to assay. At the time of the assay, media was exchanged to phenol red-free NM+Dox and supplemented with CellTiter 96 Aqueous One Solution Reagent according to the manufacturer’s instructions (Promega). The reaction was allowed to proceed for 4 h before absorbance was read at 490 nm using a microplate reader (Tecan). Media-only blanks were subtracted, and absorbance values were normalized to baseline CNO treatments, as noted. For 3D viability assays, coculture spheres containing OptoNeurons and Astrostim cells (10:1) were treated with or without CNO for varying time points and then incubated with equal volumes of phenol red–free media and CellTiter-Glo 3D Reagent (Promega) in opaque-walled, clear-bottom 96-well plates. The viability assay was carried out according to the manufacturer’s instructions and luminescence was measured using a microplate reader (Tecan). As luminescence varied linearly with sphere size, the measured values were normalized to cross-sectional areas of each sphere.
Cvetkovic C., Patel R., Shetty A., Hogan M.K., Anderson M., Basu N., Aghlara-Fotovat S., Ramesh S., Sardar D., Veiseh O., Ward M.E., Deneen B., Horner P.J, & Krencik R. (2022). Assessing Gq-GPCR–induced human astrocyte reactivity using bioengineered neural organoids. The Journal of Cell Biology, 221(4), e202107135.
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