Development and application of automatic high-resolution light microscopy for cell-based screens.
Large-scale microscopy-based screens offer compelling advantages for assessing the effects of genetic and pharmacological modulations on a wide variety of cellular features. However, development of such assays is often confronted by an apparent conflict between the need for high throughput, which usually provides limited information on a large number of samples, and a high-content approach, providing detailed information on each sample. This chapter describes a novel high-resolution screening (HRS) platform that is able to acquire large sets of data at a high rate and light microscope resolution using specific "reporter cells," cultured in multiwell plates. To harvest extensive morphological and molecular information in these automated screens, we have constructed a general analysis pipeline that is capable of assigning scores to multiparameter-based comparisons between treated cells and controls. This chapter demonstrates the structure of this system and its application for several research projects, including screening of chemical compound libraries for their effect on cell adhesion, discovery of novel cytoskeletal genes, discovery of cell migration-related genes, and a siRNA screen for perturbation of cell adhesion.
Paran
Yael
Y
Department of Molecular Cell Biology, Weizmann Institute of Science, Rehovot, Israel.
Lavelin
Irena
I
Naffar-Abu-Amara
Suha
S
Winograd-Katz
Sabina
S
Liron
Yuvalal
Y
Geiger
Benjamin
B
Kam
Zvi
Z
D001331:Automation; D002448:Cell Adhesion; D002460:Cell Line; D002465:Cell Movement; D020650:Combinatorial Chemistry Techniques; D019295:Computational Biology / Q000379:methods*; D003599:Cytoskeleton / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007091:Image Processing, Computer-Assisted; D007834:Lasers; D008853:Microscopy / Q000379:methods*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000378:metabolism; D012984:Software
D034741:RNA, Small Interfering