Table 2.

Summary of the multiplexed microscopy based assay readouts.

Readouts employed in this study to evaluate cell proliferation, TE, Hsp10 and Hsp70 induction and cell morphology from the same well. Whole well imaging at 4X objective magnification was conducted for the nuclei count TE readouts. Four fields per well covering about 10% of the well were imaged at 40X objective magnification to quantify cell stress as measured by Hsp10 and Hsp70 induction and to evaluate cell morphology by actin staining.

Channel	Fluorophore	Quantification	Readout	Objective magnification	384-well coverage
DAPI	Hoechst	Nuclei count	Cell proliferation	4X/40X	Whole well/10%
FITC	GFP	% GFP expressing cells	Transfection efficiency	4X/40X	Whole well/10%
Cy5	Anti-Hsp/AF633	Intensity per cell	Hsp10/70 induction	40X	10%
DsRed	Phalloidin-rhodamine	N/A	Cell morphology	40X	10%