An Automated High-throughput Array Microscope for Cancer Cell Mechanics.
Changes in cellular mechanical properties correlate with the progression of metastatic cancer along the epithelial-to-mesenchymal transition (EMT). Few high-throughput methodologies exist that measure cell compliance, which can be used to understand the impact of genetic alterations or to screen the efficacy of chemotherapeutic agents. We have developed a novel array high-throughput microscope (AHTM) system that combines the convenience of the standard 96-well plate with the ability to image cultured cells and membrane-bound microbeads in twelve independently-focusing channels simultaneously, visiting all wells in eight steps. We use the AHTM and passive bead rheology techniques to determine the relative compliance of human pancreatic ductal epithelial (HPDE) cells, h-TERT transformed HPDE cells (HPNE), and four gain-of-function constructs related to EMT. The AHTM found HPNE, H-ras, Myr-AKT, and Bcl2 transfected cells more compliant relative to controls, consistent with parallel tests using atomic force microscopy and invasion assays, proving the AHTM capable of screening for changes in mechanical phenotype.
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Jeremy A
JA
Cribb
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Lukas D
LD
Osborne
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Kellie
K
Beicker
Department of Biostatistics, UNC-Chapel Hill, Chapel Hill, NC United States of America.
Matthew
M
Psioda
Department of Medicine and Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina, United States of America.
Jian
J
Chen
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
E Timothy
ET
O'Brien
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.|Department of Computer Science, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Russell M
RM
Taylor Ii
Department of Computer Science, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Leandra
L
Vicci
Department of Computer Science, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Joe Ping-Lin
JP
Hsiao
Department of Computer Science, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Chong
C
Shao
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Michael
M
Falvo
Department of Biostatistics, The Lineberger Comprehensive Cancer Center, The University of North Carolina School of Medicine, Chapel Hill, North Carolina, United States of America.
Joseph G
JG
Ibrahim
Department of Pharmacology and Cancer Biology, Duke University, 450 Research Drive, Durham, NC 27710, United States of America.
Kris C
KC
Wood
Department of Medicine and Pharmacology and Cancer Biology, Duke University Medical Center, Durham, North Carolina, United States of America.
Gerard C
GC
Blobe
Department of Physics and Astronomy, UNC-Chapel Hill, Chapel Hill, NC, United States of America.
Richard
R
Superfine
D001331:Automation; D059014:High-Throughput Nucleotide Sequencing / Q000379:methods*; D006801:Humans; D008853:Microscopy / Q000295:instrumentation*; D010190:Pancreatic Neoplasms / Q000473:pathology*; D014407:Tumor Cells, Cultured