title: Combining optical tweezers and patch clamp for studies of cell membrane electromechanics.
abstract: We have designed and implemented a novel experimental setup which combines optical tweezers with patch-clamp apparatus to investigate the electromechanical properties of cellular plasma membranes. In this system, optical tweezers provide measurement of forces at piconewton scale, and the patch-clamp technique allows control of the cell transmembrane potential. A micron-size bead trapped by the optical tweezers is brought in contact with the membrane of a voltage-clamped cell, and subsequently moved away to form a plasma membrane tether. Bead displacement from the trapping center is monitored by a quadrant photodetector for dynamic measurements of tether force. Fluorescent beads and the corresponding fluorescence imaging optics are used to eliminate the shadow of the cell projected on the quadrant photodetector. Salient information associated with the mechanical properties of the membrane tether can thus be obtained. A unique feature of this setup is that the patch-clamp headstage and the manipulator for the recording pipette are mounted on a piezoelectric stage, preventing relative movements between the cell and the patch pipette during the process of tether pulling. Tethers can be pulled from the cell membrane at different holding potentials, and the tether force response can be measured while changing transmembrane potential. Experimental results from mammalian cochlear outer hair cells and human embryonic kidney cells are presented.
authors.affiliation: Department of Bioengineering, Rice University, Houston, Texas.
authors.forename: Feng
authors.initials: F
authors.lastname: Qian
authors.forename: Sergey
authors.initials: S
authors.lastname: Ermilov
authors.forename: David
authors.initials: D
authors.lastname: Murdock
authors.forename: William E
authors.initials: WE
authors.lastname: Brownell
authors.forename: Bahman
authors.initials: B
authors.lastname: Anvari