Translate this page to:
Choose Language…
In JoVE (1)
Other Publications (4)
Articles by Claire Simonnet in JoVE
JoVE General
Localized RNAi and Ectopic Gene Expression in the Medicinal Leech
1Division of Biological Sciences, University of California San Diego - UCSD, 2Department of Physics, University of California San Diego - UCSD
In this video, we show a procedure for an accurate biolistic delivery of reagents into live tissue with a novel miniature gene gun. We are knocking down the expression of the axon guidance molecule Netrin in leech embryos by delivering molecules of dsRNA into the ventral body wall and ganglia of single segments.
Other articles by Claire Simonnet on PubMed
Chaotic Mixing in a Steady Flow in a Microchannel
We report experiments on mixing of a passively advected fluorescent dye in a low Reynolds number flow in a microscopic channel. The channel is a chain of repeating segments with a custom designed profile that generates a steady three-dimensional flow with stretching and folding, and chaotic mixing. A few statistical characteristics of mixing in the flow are studied and are all found to agree with theoretical and experimental results for the flows in the Batchelor regime of mixing that are chaotic in time. The proposed microchannel provides fast and efficient mixing and is simple to fabricate.
Microtargeted Gene Silencing and Ectopic Expression in Live Embryos Using Biolistic Delivery with a Pneumatic Capillary Gun
High-throughput and High-resolution Flow Cytometry in Molded Microfluidic Devices
We describe the design, fabrication, and operation of two types of flow cytometers based on microfluidic devices made of a single cast of poly(dimethylsiloxane). The stream of particles or cells injected into the devices is hydrodynamically focused in both transverse and lateral directions, has a uniform velocity, and has adjustable diameter and shape. The cytometry system built around the first microfluidic device has fluorescence detection accuracy comparable with that of a commercial flow cytometer and can analyze as many as 17 000 particles/s. This high-throughput microfluidic device could be used in inexpensive stand-alone cytometers or as a part of integrated microanalysis systems. In the second device, a stream of particles is focused to a flow layer of a submicrometer thickness that allows imaging the particles with a high numerical aperture microscope objective. To take long-exposure, low-light fluorescence images of live cells, the device is placed on a moving stage, which accurately balances the translational motion of particles in the flow. The achieved resolution is comparable to that of still micrographs. This high-resolution device could be used for analysis of morphology and fluorescence distribution in cells in continuous flow.
Ultrafast Microfluidic Mixer with Three-dimensional Flow Focusing for Studies of Biochemical Kinetics
Studies of the kinetics of biochemical reactions, especially of folding of proteins and RNA, are important for understanding the function of biomolecules and processes in live cells. Many biochemical reactions occur rapidly and thus need to be triggered on very short time scales for their kinetics to be studied, which is often accomplished by mixing in a turbulent flow. More rapid and sample-efficient mixing is achieved in laminar flow in a microfluidic device, in which the sample is two-dimensionally (2D) focused to a thin sheet. Here we describe the design and operation of an ultrafast microfluidic mixer with three-dimensional (3D) flow focusing. The confinement of a 3D-focused sample to a narrow stream near the middle of a microchannel renders its velocity nearly uniform and makes it possible to monitor the reaction kinetics without exclusion of any parts of the sample. Hence, the sample consumption is substantially reduced and the fluorescence of the sample can be monitored without a confocal setup. Moreover, the 3D-focusing allows facile measurements of velocity of the sample with a high spatial resolution using a specially developed technique based on epi-fluorescence imaging. The data on the velocity vs. position are used to precisely calibrate the conversion between position and the reaction time, which is essential for accurate kinetic measurements. The device performs mixing on a 10 micros scale, which is comparable to that of the laminar mixers with 2D focusing. Unlike previous ultrafast laminar mixers, which were machined in hard materials, the present microfluidic device is made of a single cast of poly(dimethylsiloxane), PDMS, and is thus simpler and less expensive to manufacture.
