کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8420463 | 1545896 | 2018 | 7 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Use of photoswitchable fluorescent proteins for droplet-based microfluidic screening
ترجمه فارسی عنوان
استفاده از پروتئین های فلورسنت عکسبرداری برای غربالگری میکروفیلیدهای مبتنی بر قطره
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کلمات کلیدی
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیوتکنولوژی یا زیستفناوری
چکیده انگلیسی
Application of droplet-based microfluidics for the screening of microbial libraries is one of the important ongoing developments in functional genomics/metagenomics. In this article, we propose a new method that can be employed for high-throughput profiling of cell growth. It consists of light-driven labelling droplets that contain growing cells directly in a microfluidics observation chamber, followed by recovery of the labelled cells. This method is based on intracellular expression of green-to-red switchable fluorescent proteins. The proof of concept is established here for two commonly used biological models, E. coli and S. cerevisiae. Growth of cells in droplets was monitored under a microscope and, depending on the targeted phenotype, the fluorescence of selected droplets was switched from a “green” to a “red” state. Red fluorescent cells from labelled droplets were then successfully detected, sorted with the Fluorescence Activated Cell Sorting machine and recovered. Finally, the application of this method for different kind of screenings, in particular of metagenomic libraries, is discussed and this idea is validated by the analysis of a model mini-library.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Microbiological Methods - Volume 147, April 2018, Pages 59-65
Journal: Journal of Microbiological Methods - Volume 147, April 2018, Pages 59-65
نویسندگان
Adilya Dagkesamanskaya, Krzysztof Langer, Alexandra S. Tauzin, Catherine Rouzeau, Delphine Lestrade, Gabrielle Potocki-Veronese, Laurent Boitard, Jérôme Bibette, Jean Baudry, Denis Pompon, Véronique Anton-Leberre,