Researchers identify 10 pesticides toxic to neurons involved in Parkinson's NatureComms
. iPSC cultures were maintained in growth media . On day 0 of differentiation, confluent iPSC cultures on matrigel or geltrex were dissociated into single cells using Accutase incubated for 5–7 min at 37 degrees following a 0.5 mM EDTA in PBS wash. Accutase reaction was quenched with growth media and single cells were centrifuged, resuspended in growth media supplemented with Y-27632 at 10 µM and FGF-2 at 20 nM.
. A second analysis was then designed to count all cells brighter than three standard deviations above the average fluorescence intensity in the control wells and measure neurites. These objects were counted as THtdTomato positive cells for analysis. Built in neurite detection algorithms were applied in order to identify neurites based on THtdTomato signal in these cellular processes using the Find Neurites, CSIRO neurite analysis method.
to improve accuracy of pipetting and permit a parallel workflow to the for dispensing accuracy.
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