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",
    publisher = "Wiley-VCH Verlag",
    number = "7",

    }

    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 -