Fast Benchtop Fabrication of Laminar Flow Chambers for Advanced Microscopy Techniques

Abstract

BackgroundFluid handling technology is acquiring an ever more prominent place in laboratory science whether it is in simple buffer exchange systems, perfusion chambers, or advanced microfluidic devices. Many of these applications remain the providence of laboratories at large institutions with a great deal of expertise and specialized equipment. Even with the expansion of these techniques, limitations remain that frequently prevent the coupling of controlled fluid flow with other technologies, such as coupling microfluidics and high-resolution position and force measurements by optical trapping microscopy.MethodHere we present a method for fabrication of multiple-input laminar flow devices that are optically clear [glass] on each face, chemically inert, reusable, inexpensive, and can be fabricated on the benchtop in approximately one hour. Further these devices are designed to allow flow regulation by a simple gravity method thus requiring no specialized equipment to drive flow. Here we use these devices to perform total internal reflection fluorescence microscopy measurements as well as position sensitive optical trapping experiments.SignificanceFlow chamber technology needs to be more accessible to the general scientific community. The method presented here is versatile and robust. These devices use standard slides and coverslips making them compatible with nearly all types and models of light microscopes. These devices meet the needs of groups doing advanced optical trapping experiments, but could also be adapted by nearly any lab that has a function for solution flow coupled with microscopy.

High-Throughput Universal DNA Curtain Arrays for Single-Molecule Fluorescence Imaging

“…To further extend the utility of our wide-field surface patterning strategy, we integrated DNA curtains with a two-lane microfluidic device (Figure ). Multichannel microfluidic devices can be used to simultaneously observe enzyme function on different substrates or solution conditions. − As a proof of principle, we exploited the large number of UV-patterned DNA curtain arrays to construct a dual-lane flowcell with two distinct DNA substrates in each of the two fluidically independent lanes (Figure A, bottom panel). Biotinylated lipid bilayers were deposited concurrently in both channels by flowing all reagents through the single flow port located at the bottom of the Y-shaped flowcell (Figure S4).…”

Actin Cross-link Assembly and Disassembly Mechanics for α-Actinin and Fascin

“…Actin geometries of desired specifications were arranged using an optical trapping microscope coupled to a flow chamber (Fig. 1C) (45). Once the desired geometries (Fig.…”

“…All experiments were performed on a home-built optical trapping and multicolor fluorescence microscope. Four-input laminar flow chambers coupled with fluid reservoirs and valves were used in all trapping assays (45). All solutions were prepared in AB.…”

Microfluidic flow-cell with passive flow control for microscopy applications

“…In particular, in vitro assays such as single molecule studies require techniques to exchange buffered solutions under controlled flow rates. A wide range of approaches have been developed for fabricating fluidic devices which can perform this goal varying from high-precision, lithographically designed PDMS microfluidics or glass micro-machined devices to flow-cells formed simply by gluing together coverslips using parafilm [1][2][3][4][5][6][7][8][9][10]. A thin flow-cell geometry is additionally needed for applications in which high numerical aperture (NA) optics are combined with experimental formats that require several components to be placed in close proximity to the sample.…”