High yield, reproducible and quasi-automated bilayer formation in a microfluidic format

Research output: Contribution to journalArticleAcademicpeer-review

23 Citations (Scopus)

Abstract

A microfluidic platform is reported for various experimentation schemes on cell membrane models and membrane proteins using a combination of electrical and optical measurements, including confocal microscopy. Bilayer lipid membranes (BLMs) are prepared in the device upon spontaneous and instantaneous thinning of the lipid solution in a 100-m dry-etched aperture in a 12.5-m thick Teflon foil. Using this quasi-automated approach, a remarkable 100% membrane formation yield is reached (including reflushing in 4% of the cases), and BLMs are stable for up to 36 h. Furthermore, the potential of this platform is demonstrated for (i) the in-depth characterization of BLMs comprising both synthetic and natural lipids (1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and L--phosphatidylcholine (L--PC)/cholesterol, respectively) in terms of seal resistance, capacitance, surface area, specific capacitance, and membrane hydrophobic thickness; (ii) confocal microscopy imaging of phase separation in sphingomyelin/L--PC/cholesterol ternary membranes; (iii) electrical measurements of individual nanopores (-hemolysin, gramicidin); and (iv) indirect assessment of the alteration of membrane properties upon exposure to chemical stimuli using the natural nanopore gramicidin as a sensor
Original languageUndefined
Pages (from-to)1076-1085
Number of pages10
JournalSmall
Volume9
Issue number7
DOIs
Publication statusPublished - 8 Apr 2013

Keywords

  • natural nanopores
  • EWI-23776
  • Confocal Microscopy
  • METIS-300046
  • IR-87418
  • Membranes
  • Micro-fluidics
  • BLM

Cite this

@article{b81179c770f1453d96ec46bd1a181132,
title = "High yield, reproducible and quasi-automated bilayer formation in a microfluidic format",
abstract = "A microfluidic platform is reported for various experimentation schemes on cell membrane models and membrane proteins using a combination of electrical and optical measurements, including confocal microscopy. Bilayer lipid membranes (BLMs) are prepared in the device upon spontaneous and instantaneous thinning of the lipid solution in a 100-m dry-etched aperture in a 12.5-m thick Teflon foil. Using this quasi-automated approach, a remarkable 100{\%} membrane formation yield is reached (including reflushing in 4{\%} of the cases), and BLMs are stable for up to 36 h. Furthermore, the potential of this platform is demonstrated for (i) the in-depth characterization of BLMs comprising both synthetic and natural lipids (1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and L--phosphatidylcholine (L--PC)/cholesterol, respectively) in terms of seal resistance, capacitance, surface area, specific capacitance, and membrane hydrophobic thickness; (ii) confocal microscopy imaging of phase separation in sphingomyelin/L--PC/cholesterol ternary membranes; (iii) electrical measurements of individual nanopores (-hemolysin, gramicidin); and (iv) indirect assessment of the alteration of membrane properties upon exposure to chemical stimuli using the natural nanopore gramicidin as a sensor",
keywords = "natural nanopores, EWI-23776, Confocal Microscopy, METIS-300046, IR-87418, Membranes, Micro-fluidics, BLM",
author = "{Schulze Greiving-Stimberg}, {Verena Carolin} and Bomer, {Johan G.} and {van Uitert}, I. and {van den Berg}, Albert and {le Gac}, Severine",
note = "eemcs-eprint-23776",
year = "2013",
month = "4",
day = "8",
doi = "10.1002/smll.201201821",
language = "Undefined",
volume = "9",
pages = "1076--1085",
journal = "Small",
issn = "1613-6810",
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High yield, reproducible and quasi-automated bilayer formation in a microfluidic format. / Schulze Greiving-Stimberg, Verena Carolin; Bomer, Johan G.; van Uitert, I.; van den Berg, Albert; le Gac, Severine.

In: Small, Vol. 9, No. 7, 08.04.2013, p. 1076-1085.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - High yield, reproducible and quasi-automated bilayer formation in a microfluidic format

AU - Schulze Greiving-Stimberg, Verena Carolin

AU - Bomer, Johan G.

AU - van Uitert, I.

AU - van den Berg, Albert

AU - le Gac, Severine

N1 - eemcs-eprint-23776

PY - 2013/4/8

Y1 - 2013/4/8

N2 - A microfluidic platform is reported for various experimentation schemes on cell membrane models and membrane proteins using a combination of electrical and optical measurements, including confocal microscopy. Bilayer lipid membranes (BLMs) are prepared in the device upon spontaneous and instantaneous thinning of the lipid solution in a 100-m dry-etched aperture in a 12.5-m thick Teflon foil. Using this quasi-automated approach, a remarkable 100% membrane formation yield is reached (including reflushing in 4% of the cases), and BLMs are stable for up to 36 h. Furthermore, the potential of this platform is demonstrated for (i) the in-depth characterization of BLMs comprising both synthetic and natural lipids (1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and L--phosphatidylcholine (L--PC)/cholesterol, respectively) in terms of seal resistance, capacitance, surface area, specific capacitance, and membrane hydrophobic thickness; (ii) confocal microscopy imaging of phase separation in sphingomyelin/L--PC/cholesterol ternary membranes; (iii) electrical measurements of individual nanopores (-hemolysin, gramicidin); and (iv) indirect assessment of the alteration of membrane properties upon exposure to chemical stimuli using the natural nanopore gramicidin as a sensor

AB - A microfluidic platform is reported for various experimentation schemes on cell membrane models and membrane proteins using a combination of electrical and optical measurements, including confocal microscopy. Bilayer lipid membranes (BLMs) are prepared in the device upon spontaneous and instantaneous thinning of the lipid solution in a 100-m dry-etched aperture in a 12.5-m thick Teflon foil. Using this quasi-automated approach, a remarkable 100% membrane formation yield is reached (including reflushing in 4% of the cases), and BLMs are stable for up to 36 h. Furthermore, the potential of this platform is demonstrated for (i) the in-depth characterization of BLMs comprising both synthetic and natural lipids (1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) and L--phosphatidylcholine (L--PC)/cholesterol, respectively) in terms of seal resistance, capacitance, surface area, specific capacitance, and membrane hydrophobic thickness; (ii) confocal microscopy imaging of phase separation in sphingomyelin/L--PC/cholesterol ternary membranes; (iii) electrical measurements of individual nanopores (-hemolysin, gramicidin); and (iv) indirect assessment of the alteration of membrane properties upon exposure to chemical stimuli using the natural nanopore gramicidin as a sensor

KW - natural nanopores

KW - EWI-23776

KW - Confocal Microscopy

KW - METIS-300046

KW - IR-87418

KW - Membranes

KW - Micro-fluidics

KW - BLM

U2 - 10.1002/smll.201201821

DO - 10.1002/smll.201201821

M3 - Article

VL - 9

SP - 1076

EP - 1085

JO - Small

JF - Small

SN - 1613-6810

IS - 7

ER -