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Thermo multiskan spectrum

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The Thermo Multiskan Spectrum is a multi-mode microplate reader capable of absorbance, fluorescence, and luminescence measurements. It is designed to support a wide range of assays and applications in a laboratory setting.

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Lab products found in correlation

Mtt solution, by Thermo Fisher Scientific (2 mentions) Methyl thiazolyl tetrazolium (mtt), by Merck Group (2 mentions) Methyl thiazolyl tetrazolium (mtt), by Avantor (1 mentions) Methyl thiazolyl tetrazolium (mtt), by Thermo Fisher Scientific (1 mentions) Cck 8 reagent, by Dojindo Laboratories (1 mentions) Griess reagent, by Merck Group (1 mentions) Prismpad, by GraphPad (1 mentions)

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Multiskan Spectrum (484 citations) Thermo Multiskan Spectrum spectrophotometer (15 citations) Thermo Scientific Multiskan Spectrum (9 citations) Thermo Multiskan Spectrum plate reader (6 citations) Thermo Multiskan Spectrum Reader (3 citations) Multiskan Spectrum Thermo Electron Corporation reader (2 citations) Thermo Multiskan™ Spectrum microplate reader (2 citations)

15 protocols using thermo multiskan spectrum

1

Ursolic Acid Cytotoxicity Evaluation

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Cell viability was assessed using the MTT assay. SK-Hep-1 cells were plated at a density of 2 × 104 cells/well in 96-well plates and incubated overnight. After removing the MEM-α medium, the cells were treated with various concentrations (10, 20, 30, 40, 50, and 60 μM) of UA for 24, 48, and 72 h. After treatment, cells were treated with MTT (1 mg/mL) and incubated for 2 h at 37 °C. The medium was removed and the purple-blue MTT formazan precipitate was dissolved in 100 μL of DMSO. Absorbance was measured at 590 nm in a microplate reader (Thermo Multiskan SPECTRUM, Thermo Fisher Scientific, Waltham, MA, USA). Results are expressed as a percentage of the untreated controls. The rate of proliferation was calculated with the following formula:

where ODtest and ODblank are the optical density of the test substances and the blank controls, respectively.
Chuang W.L., Lin P.Y., Lin H.C, & Chen Y.L. (2016). The Apoptotic Effect of Ursolic Acid on SK-Hep-1 Cells is Regulated by the PI3K/Akt, p38 and JNK MAPK Signaling Pathways. Molecules, 21(4), 460.
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2

Cell Proliferation Assay of Wild-type and Mutant Cell Lines

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The cell proliferation of wild-type cell lines (HCEC, HCT116 and SW480) and the mutant cell lines (HCEC/Y641F, HCEC/Y641N, HCEC/Y641S, HCEC/Y641C and HCEC/Y641H) were assessed by the widely used MTT assay. In brief, the cells in the logarithmic growth phase were harvested and the cell density was adjusted to ~5×104/ml. A total of 100 µl cell suspension for each cell line was seeded onto 96-well plates with a final cell density of ~5,000 cells/well and cultured in the described medium and atmosphere. MTT (20 µl; Sigma-Aldrich; Merck KGaA) was added after 24, 48 and 72 h of incubation, followed by 4 h incubation in the same conditions, and the supernatant was then removed by centrifugation (1,000 × g for 10 min at room temperature). A total of 150 µl DMSO was added to each well, and the plates were oscillated at a lower speed (100 rpm) until the crystals fully dissolved. The absorbance of each well was measured at a wavelength of 570 nm using Thermo Multiskan Spectrum (Thermo Fisher Scientific, Inc.). Each experiment was repeated three times under the same conditions.
Chen Z., Yang P., Li W., He F., Wei J., Zhang T., Zhong J., Chen H, & Cao J. (2017). Expression of EZH2 is associated with poor outcome in colorectal cancer. Oncology Letters, 15(3), 2953-2961.
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3

Cell Proliferation Assay with CCK-8

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The rate of cell proliferation was assessed using the Cell Counting Kit-8 (CCK-8) purchased from Beyotime Institute of Biotechnology (Jiangsu, China), and followed the manufacturer's protocol. Cell lines transfected with shRNA or gRNA construct were seeded onto 96-well plates at a density of ~4,000 cells/well and incubated for 24, 48 and 72 h under the aforementioned culture conditions. A total of 10 µl of CCK-8 was added to each well 2 h prior to incubation completion. The absorbance of each well was measured using Thermo Multiskan® Spectrum (Thermo Fisher Scientific, Inc.) at a wavelength of 450 nm. All the experimental procedures were repeated at least three times independently.
Cai H., Xie X., Ji L., Ruan X, & Zheng Z. (2017). Sphingosine kinase 1: A novel independent prognosis biomarker in hepatocellular carcinoma. Oncology Letters, 13(4), 2316-2322.
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4

Cell Viability Assessment by MTT Assay

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Cell viability was evaluated by the 3-(4,5-dimethylthiazol-2-y1)-2,5-diphenyltetrazolium bromide (MTT) assay as previously described (20 (link)). MTT was obtained from Amresco (St. Louis, MO, USA). Briefly, MG-63 cells were seeded in 96-well plates overnight and then treated with various concentrations of DG for 24, 48 or 72 h. The optical density of the MTT formazan product was measured at a wavelength of 590 nm with a microplate reader (Thermo Multiskan SPECTRUM Thermo Fisher Scientific). Results are expressed as a percentage of the untreated controls. Data were calculated as the percentage of proliferation using the following formula: Proliferation (%) = (ODtest−ODblank) × 100, where ODtest and ODblank are the optical density (OD) of the test substances and the blank controls, respectively.
CHENG C.H., CHENG Y.P., CHANG I.L., CHEN H.Y., WU C.C, & HSIEH C.P. (2015). Dodecyl gallate induces apoptosis by upregulating the caspase-dependent apoptotic pathway and inhibiting the expression of anti-apoptotic Bcl-2 family proteins in human osteosarcoma cells. Molecular Medicine Reports, 13(2), 1495-1500.
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5

Evaluating C-PMMA and NAC-p Biocompatibility

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A total of 200 μL α-MEM with MC3T3-E1 cells at 1 × 104 cells per mL was cultivated on germ-free glass sheets in 48-well culture plates for 24 h. Then, the cell culture medium was replaced with 200 μL α-MEM, C-PMMA extracts and NAC-p (5% MDO-co-MMA) extracts as negative control, control and experimental groups, respectively. The medium was renewed every day. The cells were stained with hematoxylin and eosin (HE) after incubation for 1, 3, or 7 days. The cell state was evaluated using a light microscope at 40× magnification.
Then 100 μL of the abovementioned cell suspension was added to the 96-well culture plates. The medium was replaced with 100 μL α-MEM, C-PMMA extracts and NAC-p (5% MDO-co-MMA) extracts after 24 h as negative control, control and experimental groups, respectively. The medium was renewed every day. Then 10 μL CCK8 reagent (Dojindo, Japan) was added to the medium at 1, 3, and 5 day, and the absorbance at 450 nm (OD450) was measured at 4 h later by using a microplate reader (Thermo Multiskan Spectrum, Thermo Scientific, USA). All assays were repeated five times.
Zhao K., Pi B., Zhao L., Tian S., Ge J., Yang H., Sha W, & Wang L. (2019). Influence of N-acetyl cysteine (NAC) and 2-methylene-1,3-dioxepane (MDO) on the properties of polymethyl methacrylate (PMMA) bone cement. RSC Advances, 9(21), 11833-11841.
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6

Griess Assay for Nitric Oxide

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Nitrite in the culture supernatants was measured by the Griess reaction as an indicator of NO production. The BV2 cells were seeded in 12-well plates (1×105 cells/well), incubated overnight, and thereafter co-treated with various concentrations of HJ (100 and 500 µg/ml) and LPS (1 µg/ml) for 24 h. Following LPS stimulation, a 100 µl volume of culture supernatants from each sample were mixed with the same volume of Griess reagent (G4410; Sigma-Aldrich) and then incubated at room temperature for 15 min. The NO concentration was determined by measuring the absorbance at 540 nm on a microplate reader (Thermo Multiskan Spectrum; Thermo Fisher Scientific, Waltham, MA, USA). The nitrite concentration was calculated with reference to a sodium nitrite standard curve generated with known concentrations.
Park T.S., Ryu Y.K., Park H.Y., Kim J.Y., Go J., Noh J.R., Kim Y.H., Hwang J.H., Choi D.H., Oh W.K., Lee C.H, & Kim K.S. (2016). Humulus japonicus inhibits the progression of Alzheimer's disease in a APP/PS1 transgenic mouse model. International Journal of Molecular Medicine, 39(1), 21-30.
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7

Evaluating Nanoparticle Biocompatibility with Hemolysis

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The biocompatibility of the NPs with sheep blood cells was evaluated with a hemolysis assay. Briefly, NPs were incubated with 3% red blood cell suspensions at different concentrations (19.6, 39.2, 78.4, 156.8, 313.6, 627.2 μg/mL) for 1 h at 37 °C in 5% CO2. PBS was used as the negative control, and the positive control was a 1% w/v solution of Triton X-100. The cells were then centrifuged at 3000 rpm for 5 min, the supernatants were carefully collected and the absorbance of the supernatants was detected with a Thermo Multiskan Spectrum spectrophotometer (Thermo Scientific Inc., Waltham, MA, USA) at a wavelength of 450 nm. The hemolytic percentage (hemolysis %) was calculated according to the following equation: Hemolysis % = [A450 (NPs) − A450 (PBS)]/[A450 (1% Triton X-100) − A450 (PBS)] × 100%.
Yao P., Wang X., Wang Q., Dai Q., Peng Y., Yuan Q., Mou N., Lv S., Weng B., Wang Y, & Sun F. (2023). Cyclic RGD-Functionalized pH/ROS Dual-Responsive Nanoparticle for Targeted Breast Cancer Therapy. Pharmaceutics, 15(7), 1827.
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8

Cytotoxicity of Licochalcone A in Osteosarcoma

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The cytotoxic activity of Licochalcone A was tested using the MTT assay. The human osteosarcoma HOS and MG-63 cells were seeded in 24-well plates for overnight. The cells were treated with different concentrations of Licochalcone A for 24 h or 48 h. At the end of the assay time, the cells was incubated with 15 µL of MTT solution (5 mg/mL) (Invitrogen, Carlsbad, CA, USA) for 2 h at 37 °C. After removing the cultured medium, 200 µL of dimethyl sulfoxide (DMSO) was added to each well. Absorbance at 590 nm of the dissolved formazan product was read on an automated microplate spectrophotometer (Thermo Multiskan SPECTRUM, Thermo Fisher Scientific, Waltham, MA, USA). The half maximal inhibitory concentration (IC50) for HOS and MG-63 cells were calculated by the “Forecast” function in Microsoft Excel.
Shen T.S., Hsu Y.K., Huang Y.F., Chen H.Y., Hsieh C.P, & Chen C.L. (2019). Licochalcone A Suppresses the Proliferation of Osteosarcoma Cells through Autophagy and ATM-Chk2 Activation. Molecules, 24(13), 2435.
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9

Carnosol Cytotoxicity in Osteosarcoma

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The human osteosarcoma HOS and MG-63 cells were seeded in 24-well plates for 24 h. The cells were exposed to different concentrations of carnosol for 24 h. At the end of the assay time, 20 µL of MTT solution (5 mg/mL) (Invitrogen, Carlsbad, CA, USA) was added to each well, and then incubated for 2 h at 37 °C. After removing the cultured medium, 200 µL of dimethyl sulfoxide (DMSO) was added to each well. Absorbance at 590 nm of the dissolved formazan product was read using a spectrophotometric plate reader (Thermo Multiskan SPECTRUM, Thermo Fisher Scientific, Waltham, MA, USA). The half maximal inhibitory concentration (IC50) for HOS and MG-63 cells were calculated by the “Forecast” function in Microsoft Excel.
Hsu C.T., Huang Y.F., Hsieh C.P., Wu C.C, & Shen T.S. (2018). JNK Inactivation Induces Polyploidy and Drug-Resistance in Coronarin D-Treated Osteosarcoma Cells. Molecules : A Journal of Synthetic Chemistry and Natural Product Chemistry, 23(9), 2121.
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10

TEAC Assay for Antioxidant Capacity

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TEAC assay was performed according to Al-Duais, Müller, Böhm and Jetschke ( 2009) with some modifications. 25 mL of ABTS •+ radical cation aqueous solution (3.6 g/L) was added to 25 mL of potassium persulfate aqueous solution (0.7 g/L) and stored in the dark for 16 h at room temperature.
The solution was diluted with ethanol until reaching an absorbance at 750 nm of 0.700 (± 10%) and maintained at 30 °C. 10 µL of ethanolic extract was mixed with the prepared solution and absorbance was read at 750 nm after 5 min using a UV/VIS Thermo Multiskan Spectrum spectrophotometer (Thermo Scientific, Waltham, USA). Ethanol blanks (0.70 g/g) were run in each assay. A calibration curve was built with Trolox (0.005-0.250 mg/L). Results were expressed as mg of trolox equivalents (TE) per 100 g dm.
Plazzotta S., Ibarz R., Manzocco L, & Martín-Belloso O. (2020). Optimizing the antioxidant biocompound recovery from peach waste extraction assisted by ultrasounds or microwaves. Ultrasonics sonochemistry, 63.
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