The production of micron-size droplets using microfluidic tools offers new opportunities to carry out biological assays in a controlled environment. We apply these strategies by using a flow-focusing microfluidic device to encapsulate Xenopus egg extracts, a biological system recapitulating key events of eukaryotic cell functions in vitro. We present a method to generate monodisperse egg extract-in-oil droplets and use high-speed imaging to characterize the droplet pinch-off dynamics leading to the production of trains of droplets. We use fluorescence microscopy to show that our method does not affect the biological activity of the encapsulated egg extract by observing the self-organization of microtubules and actin filaments, two main biopolymers of the cell cytoskeleton, encapsulated in the produced droplets. We anticipate that this assay might be useful for quantitative studies of biological systems in a confined environment as well as high throughput screenings for drug discovery.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1039/c0lc00046a | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Development of translationally active cell lysates from different filamentous fungi for application in cell-free protein synthesis.
Enzyme Microb Technol
January 2025
Institute of Biotechnology, Brandenburg University of Technology Cottbus-Senftenberg, Universitätsplatz 1, Senftenberg 01968, Germany. Electronic address:
There is an enormous potential for cell-free protein synthesis (CFPS) systems based on filamentous fungi in view of their simple, fast and mostly inexpensive cultivation with high biomass space-time yields and in view of their catalytic capacity. In 12 of the 22 different filamentous fungi examined, in vitro translation of at least one of the two reporter proteins GFP and firefly luciferase was detected. The lysates showing translation of a reporter protein usually were able to synthesize a functional cell-free expressed unspecific peroxygenase (UPO) from the basidiomycete Cyclocybe (Agrocybe) aegerita.
View Article and Find Full Text PDFT-cell prolymphocytic leukemia (T-PLL) is an aggressive lymphoid malignancy with limited treatment options. To discover new treatment targets for T-PLL, we performed high-throughput drug sensitivity screening on 30 primary patient samples ex-vivo. After screening over 2'800 unique compounds, we found T-PLL to be more resistant to most drug classes, including chemotherapeutics, compared to other blood cancers.
View Article and Find Full Text PDFA Fast-Pass, Desorption Electrospray Ionization Mass Spectrometry Strategy for Untargeted Metabolic Phenotyping.
J Am Soc Mass Spectrom
January 2025
Department of Chemistry, Center for Innovative Technology, Vanderbilt University, Nashville, Tennessee 37235, United States.
Desorption electrospray ionization mass spectrometry imaging (DESI-MSI) provides direct analytical readouts of small molecules that can be used to characterize the metabolic phenotypes of genetically engineered bacteria. In an effort to accelerate the time frame associated with the screening of mutant libraries, we have developed a high-throughput DESI-MSI analytical workflow implementing a single raster line-scan strategy that facilitates the collection of location-resolved molecular information from engineered strains on a subminute time scale. Evaluation of this "Fast-Pass" DESI-MSI phenotyping workflow on analytical standards demonstrated the capability of acquiring full metabolic profiling information with a throughput of ∼40 s per sample.
View Article and Find Full Text PDFAutomated High-Throughput Affinity Capture-Mass Spectrometry Platform with Data-Independent Acquisition.
J Proteome Res
January 2025
Discovery Research, AbbVie, Inc., 1 North Waukegan Rd., North Chicago, Illinois 60064, United States.
Affinity capture (AC) combined with mass spectrometry (MS)-based proteomics is highly utilized throughout the drug discovery pipeline to determine small-molecule target selectivity and engagement. However, the tedious sample preparation steps and time-consuming MS acquisition process have limited its use in a high-throughput format. Here, we report an automated workflow employing biotinylated probes and streptavidin magnetic beads for small-molecule target enrichment in the 96-well plate format, ending with direct sampling from EvoSep Solid Phase Extraction tips for liquid chromatography (LC)-tandem mass spectrometry (MS/MS) analysis.
View Article and Find Full Text PDFQuantifying Competitive Fitness in Yeast with High-Throughput Fluorescence Microscopy Imaging.
Curr Protoc
January 2025
Department of Microbiology, University of Manitoba, Winnipeg, Manitoba, Canada.
Competitive fitness is a fundamental concept in evolutionary biology that captures the ability of organisms to survive, reproduce, and compete for resources in their environment. Competitive fitness is typically assessed in the lab by growing two or more competitors together and measuring the frequency of each at multiple time points. Traditional microbial competitive fitness assays are labor intensive and involve plating on solid medium and counting colonies.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!