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.

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Main benefits

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)

Product	Cat. #	Company	Link
Micropipette	ES-Blastocyst Injection Pipettes – Blunt	Biomedical Instruments	Biomedical Instruments
Micromanipulator (you can choose between one of the two selected micromanipulators	MMO-4 (upgraded MMO-202ND)	Narishige	Narishige
Micromanipulator (you can choose between one of the two selected micromanipulators	TransfertMan 4r	Eppendorf	Eppendorf