Micropipette aspiration package

[E-MASPI01]

    A complete setup for starting micropipette aspiration experiments

    The Fluigent micropipette aspiration package allows one to manipulate and characterize cells at the single-cell level. It is a powerful non-invasive technique to evaluate how the biomechanical properties of single cells or tissues govern cell shape, cell response to mechanic stimuli, transition from nontumorigenic to tumorigenic state, or morphogenesis.

    Main benefits
    • compact
      High sensitivity and resolution

      Pressure control

    • Time saving

      Non-invasive method

    • Cost-effective

      Easy to build and operate

    Features

    Extreme precision

    Fluigent instruments are the only products with the ability to control small pressure increments (0.007 mbar) at low pressure (0.1-10 mbar). They allow the investigation of subcellular dynamics like cytoskeleton structural and organizational modifications that are not accessible with confocal microscopy.

    Pressure control

    Manual aspiration causes inter-operator variability as the applied pressure cannot be exactly quantified. In contrast, Fluigent pressure regulators deliver the set pressure with +/- 0.1% precision.

    Engineered package

    Micropipette aspiration with Fluigent is compact, can fit any microscope and micropipette and is controlled by an intuitive software package. Competitive technologies like AFM, cytoindenter and optical tweezers are expensive, necessitate specific training and may require a dedicated microscope

    Non-invasive

    Micropipette aspiration allows for repetitive measurements on the same sample. Variations in cell tension of individual cells within a tissue can be monitored over time.

    Repeat experiments

    Cell surface tension can be measured in 3 to 5 minutes

    Related applications

    Applications

    • Cell mechanical properties measurement: Many biological processes are characterized by changes in cell stiffness: cells entering mitosis [1], tumor cells transitioning to premalignant stage [2], red blood cells infected with malaria [3]… These changes occur at cell scale and require precise measurement to accurately quantify cell stiffness. A Dual pipette aspiration assay is a tool to evaluate the relative contribution of cell-cell tension versus cell-medium tension at the cell-cell interface by separating contacting cells (Maitre et al Sciences 2012).
    • Single cell manipulation: Micropipette aspiration allows spatial positioning of single cells or clusters of cells. Single cell positioning is necessary for single cell analysis or clonal cell line development.
    • Tension heterogeneity within tissue: Evaluating cell tension at the single cell level allows for mapping the of tensions within a tissue. It is particularly useful to investigate the forces driving tissue morphogenesis or embryogenesis ( Maitre et al, 2016, Nature)
    • In vitro diagnostic: Measuring stiffness with cellular resolution can be a powerful tool to detect abnormal behaviors that are not accessible or perceptible under a microscope. As an example, mechanical properties can predict the viability of embryos within hours after fertilization even though viable and non-viable embryos are morphologically indistinguishable at this stage [4]

    Pressure are the gold standard for microaspiration

    Micropipette aspiration requires high control over the forces applied to cells. A fast response time is necessary as transitional states should be minimized.

    Only Fluigent can provide products with pressure ranges of 0 – (+/-) 25 mbar, or 0 – (+/-) 69 mbar, to apply small pressure increments (0.007 mbar) at low pressure (0.1 -10 mbar).

    The Flow EZ™ pressure controllers are particularly suited as this method requires applying forces ranging from 10pN to 1nN accurately.

    References

    [1] Théry M, Bornens M, Get round and stiff. 2008, HFSP J, 2(2):65-71.

    [2] Tavares S et al, actin stress fiber organization promotes cell stiffening and proliferation of pre-invasive breast cancer cells. 2017, Nat Commun. 8:15237.

    [3] Guo Q et al, Microfluidic biomechanical assay for red blood cells parasitized by Plasmodium falciparum. 2012, Lab Chip; 12(6):1143-50.

    [4] Yanez LZ et al, human oocyte developmental potential is predicted by mechanical properties within hours after fertilization, 2016, Nat Commun. 7:10809

    Main products of the package

    Specifications

    LineUp Flow EZ pressure controller (-25 mbar) x1
    LineUp LINK Module (software control) x1
    Fluiwell-1C-15 ml HPMFCS (15 mL fluid reservoir)
    Compact Vacuum Pump x1
    Complete Linear Stage for reservoir displacement x1
    Tubing & fitting kit (from the reservoir to the micropipette holder) x1

    Additional items required for micropipette aspiration experiments (not included in Fluigent package)

    ProductCat. #CompanyLink
    MicropipetteES-Blastocyst Injection Pipettes – BluntBiomedical InstrumentsBiomedical Instruments
    Micromanipulator (you can choose between one of the two selected micromanipulatorsMMO-4 (upgraded MMO-202ND)NarishigeNarishige
    Micromanipulator (you can choose between one of the two selected micromanipulatorsTransfertMan 4rEppendorfEppendorf

    OxyGEN

    Control in real-time, protocol automation, data record and export
    ver. 1.0.0.0 or more recent

    See the offer


    Software Development Kit

    Custom software application
    ver. 21.0.0.0 or more recent

    See the offer

    Protocol for building the micro-aspiration system and how to use it for subcellular surface tension mapping

    Guevorkian K,Maître JL.Micropipette aspiration: A unique tool for exploring cell and tissue mechanics in vivo. MethodsCellBiol. 2017;139:187-201

    Selected publications from our customers

    Maître JL et al, Asymmetric division of contractile domains couples cell positioning and fate specification, Nature. 2016 Aug 18;536(7616):344-34

    Biro M, Maître JL, Dual pipette aspiration: a unique tool for studying intercellular adhesion. Methods CellBiol. 2015;125:255-67

    Porazinski S et al, YAP is essential for tissue tension to ensure vertebrate 3D body shape. Nature. 2015 May 14;521(7551):217-221

    Maître JL et al, Pulsatile cell-autonomous contractility drives compaction in the mouse embryo. Nat Cel lBiol. 2015 Jul;17(7):849-55

    Maître JL et al, Adhesion functions in cell sorting by mechanically coupling the cortices of adhering cells. Science. 2012;338(6104):253-6