Lateral patch-clamping in a standard 1536-well microplate format.
Lateral patch-clamping has emerged as a chip-based platform for automation of the conventional patch-clamp technique, the 'gold' standard for studying cellular ion channels. The conventional technique, as it relies on skilled-maneuver of glass micropipettes to patch cells, is extremely delicate, low in throughput, and thus cannot be used for primary screening of compounds against ion channels. Direct integration of glass capillaries on silicon provides lateral junctions for automated trapping and patching of cells. We demonstrate here a method of scaling up the lateral junctions to a standard 1536-well microtiter plate format. A single unit of 1536-well plate has been formed here on a 9 mm by 9 mm microstructured silicon with the inclusive of 16 wells molded in a capping layer made of polydimethylsiloxane (PDMS). The silicon substrate provides integrated glass capillaries (total 12) and their associated microfluidic network. Each glass capillary has an independent access through a dedicated well in PDMS and leads to a centralized channel in which cell suspension can be delivered through one of the remaining 4 wells. The unit has been tested on RBL-1 cells by recording whole-cell activity from inwardly rectifying endogenous potassium channels. A revised test protocol has been prescribed to avoid inaccurate readings due to altered ionic composition of the recording buffer when a typical suction is applied to capture cells.
Tang
Kum Cheong
KC
Institute of Microelectronics (IME), 11 Science Park Road, Science Park II, Singapore.
Reboud
Julien
J
Kwok
Yuan Li
YL
Peng
Shu Ling
SL
Yobas
Levent
L
D000200:Action Potentials / Q000502:physiology*; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002460:Cell Line; D004566:Electrodes; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006801:Humans; D008564:Membrane Potentials / Q000502:physiology*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009474:Neurons / Q000502:physiology*; D018408:Patch-Clamp Techniques / Q000295:instrumentation* / Q000379:methods