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Imark microplate

Manufactured by Bio-Rad
Sourced in United States

The IMark microplate is a laboratory equipment designed for absorbance measurements in microplate format. It is used to quantify the amount of light absorbed by samples in a microplate, which can be used to measure various biological and chemical reactions.

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7 protocols using imark microplate

1

Photothermal Therapy of Melanoma Cells

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The cancer cells used in the photothermal therapy were human malignant melanoma (A375). They were obtained from the Cell Lab of the Cell Resource Center of the Chinese Academy of Sciences. The cell culture methods are very similar to those described in Reference [25 (link)].
Cell viability was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. This method is similar to those described in Reference 25. The 96-well plate was then put into an iMark Microplate (BioRad) to measure optical densities (OD) at 490 nm. The cell morphology of A375 cells incubated with GNRs at different concentrations are shown in Figure S2.
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2

Ferric Reducing Antioxidant Power Assay

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The ferric reducing antioxidant power assay followed the method of Moyo et al. [53 ]. The FRAP reagent was prepared by sequentially mixing 25 mL of acetate buffer, 2.5 mL of 2,4,6-Tripyridyl-S-triazine (TPTZ), and 2.5 mL of FeCl3·6H2O. The mixture was submerged and kept in a 37 °C water bath. Briefly, 90 µL volume of double-distilled water (ddH2O), 30 µL of a sample (1 mg/mL) was combined with 900 µL of the FRAP reagent and incubated for 30 min the dark. The resultant ferrous tripyridyltriazine-coloured complex was read at 595 nm using an absorbance reader (iMark microplate, Bio-Rad laboratories 168-1130). Results were expressed as µmol TE/g dw.
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3

Cytotoxicity Evaluation of Nanomaterials

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Determination of concentration- and time-dependent cytotoxicity was carried out using the WST-1 assay (Roche Applied Science, USA). Panc-1 cells were seeded in 96-well plates for 24 hours, after which the cells were treated with nanomaterial and/or drug. After incubation, the treatment medium was removed, and the cells were washed 3 times with 1× PBS buffer. 100 µl of fresh complete growth medium were then added to each well, followed by 10 µl of the WST-1 reagent. The plates were then incubated in the dark at 37°C. After 2 h, absorbance was measured using an iMark microplate (Bio-Rad, USA) reader at a wavelength of 450 nm. All of the control and treatment samples were run in triplicate.
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4

Evaluating NF-κB Activation by Bacterial Strains

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HEK293-Blue reporter cells stably expressing human TLR4 (HEK-TLR4), were cultured according to the manufacturer’s instructions. Briefly, HEK-TLR4 cells were maintained in DMEM 4.5 g/L D-glucose supplemented with 10% (v/v) heat-inactivated FBS, 4 mM L-Glutamine, 100 U/ml penicillin, 100 μg/ml streptomycin, 100 μg/ml normocin, 1X HEK-Blue selection media.
Briefly, cells were washed with PBS, dissociated in PBS and collected in growth media. Cells were plated in 96-well plates at a density of 25,000 cells/well. To evaluate the effect of bacteria strains on LPS inducing NF-κB promoter activation, cells were treated with 10 ng/ml LPS in presence or absence of 10% supernatants and incubated in a CO2 incubator. Treatments proceeded for 22 h at 37°C and 5% CO2, after which the detection of Secreted Embryonic Alkaline Phosphatase (SEAP) activity from cell culture supernatant was performed using QUANTI-blue solution according to manufacturer’s instructions. Briefly, 20 μl of cell-free supernatant was collected and analyzed for the presence of SEAP by mixing with 200 μl of sterile-filtered QUANTI-Blue detection media. After 2 h incubation at 37°C, optical density was measured at 655 nm on a microplate reader (iMark microplate, Bio-Rad).
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5

Quantification of Hepatic Lipid Droplets

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As indicated, cells were stained with ORO (Sanchez-Hidalgo et al., 2007 (link)) and examined under a light microscope (Nikon Eclipse TS100). ORO was quantified through colorimetry after extraction with 1 mL 100% isopropanol for 10 min, followed by gentle vibration to release ORO from cells (Lin et al., 2007 (link)). The extract from individual wells was then transferred to a 96-well plate and absorbance measured at 500 nm using Bio-Rad iMark Microplate. The quantification of ORO intensity in representative liver sections was done using NIS Element software. ORO intensity was measured by selecting the color threshold of the ORO droplet and results expressed as mean intensity/area.
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6

Cytotoxicity Assay for Osmium Complex

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A stock solution of 1 was prepared in cell culture medium (5% DMSO). Approximately 5000 A2780 human ovarian cancer cells were seeded per well in 96-well plates. The cells were pre-incubated in drug-free media at 37 °C for 48 h before adding different concentrations of 1. Cells were exposed to 1 for 24 h at 37 °C and then allowed to recover for 72 h in a drug-free medium at 37 °C. Then the supernatants were removed by suction and the cells washed with PBS. The cells were The SRB assay was used to determine cell viability. Absorbance measurements of the solubilised dye (on a BioRad iMark microplate reader using a 470 nm filter) allowed the determination of viable treated cells compared to untreated controls. IC50 values (the concentration at which 50% cell death occurs) were determined as triplicates of duplicates for each complex. ICP-OES (PerkinElmer Optima 5300 DV instrument) was used to determine [Os] of the stock solution of 1, and the IC50 value was corrected for ICP factor.
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7

Cell Proliferation Assay for DAM and DAM-NPs

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The effect of DAM and DAM-NPs on the proliferation of HCT-116 and U2OS cells was investigated using the CellTiter 96 Aqueous One Solution Cell Proliferation Assay (Promega Corporation, Madison, WI, USA). The cells were seeded at a concentration of 3,000 cells/well in 96-well tissue culture plates. The cells were then treated with 1 or 10 µM DAM or DMSO as a control, and DAM-NPs (equivalent concentration of 50 µM DAM) or DAM-free NPs as controls. At 0, 1 and 3 days after treatment (for DAM-NPs and NPs) or 0, 1 and 4 days after treatment (for DAM and DMSO), 20 µl CellTiter 96 Aqueous One Solution was added to each well. The plate was then incubated at 37°C for 1 h. The absorbance at 490 nm was determined using an iMARK™ microplate absorbance reader (Bio-Rad Laboratories, Inc., Hercules, CA, USA).
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