Micropipette cell and tissue aspiration
Micropipette aspiration is a powerful non-invasive technique to evaluate how biomechanical properties of single cells or tissue govern cell shape, cell response to mechanic stimuli, transition from nontumorigenic to tumorigenic state or morphogenesis.
The Fluigent MFCS™-EZ and Flow EZ™ pressure controllers are particularly suited for this method since it requires applying forces ranging from 10pN to 1nN accurately.
Background image courtesy of Jean-Léon Maitre (Institut Curie, France)
Non-invasive: Micropipette aspiration allows repetitive measurements on the same sample. Variations in cell tension of individual cells within a tissue can be monitored over time. This method is a powerful tool to trace the spatio temporal map of tensions inducing morphogenesis ( Maitre et al, Nat Cell Biol, 2015).
Economical, easy to use and build: Micropipette aspiration with Fluigent is compact, can fit any microscope and is piloted by an intuitive software. Competitive technologies like AFM, cytoindenter and optical tweezers are expensive, necessitate specific training and may require a dedicated microscope.
Time saving: Thanks to Fluigent’s product responsiveness, a given pressure is instantaneously applied on the cell’s surface (ms range). Cell surface tension can be measured in 3 to 5 minutes (Maitre et al, Nat Cell Biol, 2015)
No inter-operator variability: Besides requiring highly skilled people, manual aspiration causes considerable inter-operator variability as the applied pressure cannot be exactly quantified. In contrast, Fluigent pressure regulators always deliver the ordered pressure with 0.1% precision.
High sensitivity and resolution: Fluigent instruments are the only products in the market delivering small pressure increments (0.007mbar) at low pressure (0.1-10mbar). They allow investigating subcellular dynamics like cytoskeleton structural and organizational modifications that are not accessible with confocal microscopy.
Cell mechanical properties measurement: Many biological processes are characterized by changes in cell stiffness: cells entering mitosis , tumor cells transitioning to premalignant stage , red blood cells infected with malaria … These mechanical changes occur at cell scale and require precise measurement to accurately quantify cell stiffness. Dual pipette aspiration assay, the duplicated version of cell aspiration set up, is another handy tool to evaluate the relative contribution of cell-cell tension versus cell-medium tension at 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 mapping the spatial map of tensions of a tissue. It is particularly performant 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 power full tool to detect abnormal behavior that are not accessible nor perceptible under a microscope. As an example, mechanical properties can predict the viability of embryos already within hours after fertilization although viable and non-viable embryos are morphologically indistinguishable at this stage .
THE MICRO-ASPIRATION APPLICATION COMPONENTS
|MICROFLUIDIC PUMP||EZ-11000001 + EZ-|
|MICROPIPETTE||ES-Blastocyst Injection Pipettes - Blunt||Biomedical Instruments||Biomedical Instruments|
|LINEAR TRANSLATION STAGE||M-UMR8.51 « Translation stage »||Newport||Newport|
|45 « Rod »||Newport 45|
|340-RC « Clamp »||Newport 340-RC|
|M-MB-3 « Compact Magnetic Base »||Newport M-MB-3|
 Théry M, Bornens M, Get round and stiff. 2008, HFSP J, 2(2):65-71.
 Tavares S et al, actin stress fiber organization promotes cell stiffening and proliferation of pre-invasive breast cancer cells. 2017, Nat Commun. 8:15237.
 Guo Q et al, Microfluidic biomechanical assay for red blood cells parasitized by Plasmodium falciparum. 2012, Lab Chip; 12(6):1143-50.
 Yanez LZ et al, human oocyte developmental potential is predicted by mechanical properties within hours after fertilization, 2016, Nat Commun. 7:10809
Selected publications from our customers
Guevorkian K,Maître JL.Micropipette aspiration: A unique tool for exploring cell and tissue mechanics in vivo.MethodsCellBiol. 2017;139:187-201
Maître JL et al, Asymmetric division of contractile domains couples cellpositioning 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.MethodsCellBiol. 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-autonomouscontractility 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
The use of water-in-oil droplets in microfluidics in high-throughput screening is rapidly gaining acceptance. The main application areas currently involve screening cells as well as genetic material for various mutations or activity. Here the aim is to isolate single DNA molecules and analyze the enzymes and proteins resulting from their expression.
Many microfluidic applications require switching between multiple solutions (such as samples or buffers) while maintaining a constant flow-rate during the course of their experiment. In this application, one MFCS™-EZ pressure channel is connected via a manifold to ten separate vials containing different aqueous solutions.
Because CTCs detection and labelling need a very precise and smooth flow control, Fluigent MFCS™ associated with the Flow-Rate Control Module and the ESS™ platform, was the designated choice to be able to efficiently control the flows of all the solutions and samples.