Dataset Information

A microfluidic pipette array for mechanophenotyping of cancer cells and mechanical gating of mechanosensitive channels.

ABSTRACT: Micropipette aspiration measures the mechanical properties of single cells. A traditional micropipette aspiration system requires a bulky infrastructure and has a low throughput and limited potential for automation. We have developed a simple microfluidic device which is able to trap and apply pressure to single cells in designated aspiration arrays. By changing the volume flow rate using a syringe pump, we can accurately exert a pressure difference across the trapped cells for pipette aspiration. By examining cell deformation and protrusion length into the pipette under an optical microscope, several important cell mechanical properties, such as the cortical tension and the Young's modulus, can be measured quantitatively using automated image analysis. Using the microfluidic pipette array, the stiffness of breast cancer cells and healthy breast epithelial cells was measured and compared. Finally, we applied our device to examine the gating threshold of the mechanosensitive channel MscL expressed in mammalian cells. Together, the development of a microfluidic pipette array could enable rapid mechanophenotyping of individual cells and for mechanotransduction studies.

SUBMITTER: Lee LM

PROVIDER: S-EPMC4256121 | biostudies-literature | 2015 Jan

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

A microfluidic pipette array for mechanophenotyping of cancer cells and mechanical gating of mechanosensitive channels.

Lee Lap Man LM Liu Allen P AP

Lab on a chip 20150101 1

Micropipette aspiration measures the mechanical properties of single cells. A traditional micropipette aspiration system requires a bulky infrastructure and has a low throughput and limited potential for automation. We have developed a simple microfluidic device which is able to trap and apply pressure to single cells in designated aspiration arrays. By changing the volume flow rate using a syringe pump, we can accurately exert a pressure difference across the trapped cells for pipette aspiratio ...[more]

PMID: 25361042

Similar Datasets

Project description:The mechanosensitive channel of large conductance (MscL) is capable of transducing mechanical stimuli such as membrane tension into an electrochemical response. MscL provides a widely-studied model system for mechanotransduction and, more generally, for how bilayer mechanical properties regulate protein conformational changes. Much effort has been expended on the detailed experimental characterization of the molecular structure and biological function of MscL. However, despite its central significance, even basic issues such as the physiologically relevant oligomeric states and molecular structures of MscL remain a matter of debate. In particular, tetrameric, pentameric, and hexameric oligomeric states of MscL have been proposed, together with a range of detailed molecular structures of MscL in the closed and open channel states. Previous theoretical work has shown that the basic phenomenology of MscL gating can be understood using an elastic model describing the energetic cost of the thickness deformations induced by MscL in the surrounding lipid bilayer. Here, we generalize this elastic model to account for the proposed oligomeric states and hydrophobic shapes of MscL. We find that the oligomeric state and hydrophobic shape of MscL are reflected in the energetic cost of lipid bilayer deformations. We make quantitative predictions pertaining to the gating characteristics associated with various structural models of MscL and, in particular, show that different oligomeric states and hydrophobic shapes of MscL yield distinct membrane contributions to the gating energy and gating tension. Thus, the functional properties of MscL provide a signature of the oligomeric state and hydrophobic shape of MscL. Our results suggest that, in addition to the hydrophobic mismatch between membrane proteins and the surrounding lipid bilayer, the symmetry and shape of the hydrophobic surfaces of membrane proteins play an important role in the regulation of protein function by bilayer membranes.

Dataset Information

A microfluidic pipette array for mechanophenotyping of cancer cells and mechanical gating of mechanosensitive channels.

Publications

A microfluidic pipette array for mechanophenotyping of cancer cells and mechanical gating of mechanosensitive channels.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets