Infinite 200 pro multifunctional microplate reader

To determine the intracellular uptake, antisarAbs were fluorescently labeled with CdTe quantum dots (QDs) through a synthetic heptapeptide HWR linker prior to encapsulation, as described in our previously published studies11 44 (link). PC-3 cells were seeded onto a plate at a density of 2.0 × 10⁵cells/well and were incubated for 24 h. Microscopy studies were performed using an Olympus IX 71S8F-3 inverted fluorescence microscope (Olympus, Tokyo, Japan) equipped with an HBO 50 W mercury arc lamp (Osram GmbH, Munich, Germany). The 545 and 580 nm excitation filters and the 610 nm emission filter were utilized. Images were acquired with an Olympus DP73 camera and processed using Stream Basic 1.7 software (Olympus) at a resolution of 1,600 × 1,200 pixels. To evaluate the competitive inhibitory effects of FA, various FA concentrations (final FA concentrations 2; 8 and 20 mM) were utilized for pre-treatment, followed by subsequent treatment with 10 μM QD-labeled antisarAbs@LIP and incubation for 4 h. The cells were then washed with medium, trypsinized and harvested. The median QD fluorescence values were analyzed using a Tecan Infinite 200 PRO multifunctional microplate reader (Tecan, Maennedorf, Switzerland) using λ_ex = 500 nm and λ_em = 610 nm.

Based on the method described by Chen et al. (2017) (link), the changes in intracellular ATP concentrations of L. monocytogenes CMCC 54004 after treatments with OOPE was measured. The final L. monocytogenes CMCC 54004 cell concentration was diluted to be 10⁸ CFU/ml with NS. The ATP assay kit (Beyotime Bioengineering Institute, Shanghai, China) was used to determine intracellular ATP concentration, and all processes were operated on an ice box. Two milliliters of cell suspensions containing different concentrations of OOPE (0 MIC, 1/4 MIC, 1/2 MIC, 1 MIC, and 2 MIC) was incubated at 37°C for 30 min and then mixed with lysis solution. The supernatant was obtained after centrifugation at 11,269 × g for 5 min and stored in an ice box to prevent the loss of ATP. Both ATP test solution and supernatant were added to a colorless transparent 96-well plate (Corning Institute, USA) to determine intracellular ATP concentration with an Infinite 200 PRO multifunctional microplate reader (Tecan, Grodlg, Austria).