352 multiskan ms microplate reader

The cellular supernatant was analyzed for the enzyme lactate dehydrogenase (LDH) using a Pierce™ LDH Cytotoxicity Assay Kit (Thermo Fisher Scientific). Lung cells in the incubator and also from the GIVES were analyzed in triplicate and scanned for measuring the absorbance at 492 and 690 nm wavelengths using Thermo/LabSystems 352 Multiskan MS Microplate Reader. The outliers were identified as those with <5% probability of occurrence relative to a normal distribution.19 LDH data were determined for each of the six wells in the exposed plate from GIVES and for the six wells in the plate in the incubator exposed to clean air. Fold change in LDH was calculated by dividing the six well mean values of the exposed samples in the GIVES by the six well mean values of the samples from the incubator. Percent death was calculated by normalizing to 100% cytotoxic conditions of a 7.5-fold change.20 Data were analyzed using an unpaired Student’s t-test where differences were considered statistically significant if P ≤ 0.05.