Nuclear morphology and cell number were convenient indices of apoptosis and cell viability. Here, we outline our implementation of image analysis strategies and statistical tools to distill large high-content data sets. Specifically, we use a B-score transformation to effectively mitigate the systematic row and column artifacts that are commonly observed during high-throughput screens of live cells. Our results demonstrate the feasibility of multi-parameter. We identify novel compounds that can significantly increase or decrease insulin gene expression. In this report, we present a novel imaging-based screening platform designed to identify compounds capable of altering the differentiation state, survival and/or proliferation of pancreatic beta-cells. To the best of our knowledge, this is the first implementation of multi-parameter, high-content, high-throughput screening in living pancreatic beta-cells. We also describe data analysis and statistical approaches that eliminate spurious results arising from row and column biases that are systematic in multiwell live cell analysis. We present results from SB431542 ALK inhibitor multiple replicate screens that highlight several extracts capable of significantly altering insulin and pdx1 promoter activity in beta-cells. We identified the active insulin gene expression promoting component, Bivittoside D, from one of these extracts. The key elements in successful high-throughput screening programs are the judicious choice of validated molecular targets and access to the greatest number and structural diversity of compounds. Roughly four decades of investigations into natural products derived from marine invertebrates have shown that they are an exceptionally rich source of novel chemotypes that frequently exhibit potent biological activities. Marine sponge extracts also capture some of the chemical diversity of the associated microorganisms. Sponges are a rich source of terpenoids, peptides, alkaloids, polyketides, glycolipids, steroids, and many compounds with mixed biogenetic origins. Notably, this library of compounds has already produced several drug candidates for cancer therapy. In the present study, the use of multiple parameters was extremely valuable, as it allowed us to eliminate many hits from compounds that were also cytotoxic and would therefore be unsuitable leads for improving beta-cell function. The resulting seven lead extracts showed no effect on cell number or nuclear staining intensity in the primary screen. In the future, it might also be useful to investigate other methods of hit selection, including those that simultaneously weigh effects using multiple parameters. From the selected seven extracts detected in the primary screen, we followed up on two extracts that had a positive and negative effect on insulin expression. From our secondary screens and follow-up studies, Bivittoside D was identified as a positive regulator of insulin gene expression. Bivittoside D is a lanostane triterpenoid with six monosaccharide units.