Abstract

Microfluidic devices allow assays to be performed using minute amounts of sample and have recently been used to control the microenvironment of cells. Microfluidics is commonly associated with closed microchannels which limit their use to samples that can be introduced, and cultured in the case of cells, within a confined volume. On the other hand, micropipetting system have been used to locally perfuse cells and surfaces, notably using push-pull setups where one pipette acts as source and the other one as sink, but the confinement of the flow is difficult in three dimensions. Furthermore, pipettes are fragile and difficult to position and hence are used in static configuration only.The microfluidic probe (MFP) circumvents the constraints imposed by the construction of closed microfluidic channels and instead of enclosing the sample into the microfluidic system, the microfluidic flow can be directly delivered onto the sample, and scanned across the sample, using the MFP. . The injection and aspiration openings are located within a few tens of micrometers of one another so that a microjet injected into the gap is confined by the hydrodynamic forces of the surrounding liquid and entirely aspirated back into the other opening. The microjet can be flushed across the substrate surface and provides a precise tool for localized deposition/delivery of reagents which can be used over large areas by scanning the probe across the surface. In this video we present the microfluidic probe1 (MFP). We explain in detail how to assemble the MFP, mount it atop an inverted microscope, and align it relative to the substrate surface, and finally show how to use it to process a substrate surface immersed in a buffer.

Highlights

  • Assembly of the microfluidic probe (MFP)1. Gas-tight glass syringes are filled with the appropriate reagents using plastic syringes and needles to ensure that no air bubbles are present

  • Microfluidic devices allow assays to be performed using minute amounts of sample and have recently been used to control the microenvironment of cells

  • The microfluidic probe (MFP) circumvents the constraints imposed by the construction of closed microfluidic channels and instead of enclosing the sample into the microfluidic system, the microfluidic flow can be directly delivered onto the sample, and scanned across the sample, using the MFP

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Summary

Assembly of the MFP

1. Gas-tight glass syringes are filled with the appropriate reagents using plastic syringes and needles to ensure that no air bubbles are present. We use a 1 -10 microliter syringe for injection, and a syringe with 5-10 times larger volume for aspiration. 2. The syringes are connected to capillary tubing using Nanotight fittings with low dead volume. 3. Capillaries are filled and checked for bubbles under the microscope. 4. The MFP chip is prefilled with buffer solution to prevent trapping of bubble when connecting the capillaries

Set-up of the MFP
Operation of the MFP
Discussion
Full Text
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