A Wide-Field Fluorescence Microscope Extension for Ultrafast Screening of One-Bead One-Compound Libraries Using a Spectral Image Subtraction Approach
| العنوان: | A Wide-Field Fluorescence Microscope Extension for Ultrafast Screening of One-Bead One-Compound Libraries Using a Spectral Image Subtraction Approach |
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| المؤلفون: | Beat Ludin, Thomas Weidemann, Martin Hintersteiner, Manfred Auer, Nhan T. Pham, Wolf Heusermann |
| المصدر: | ACS Combinatorial Science. 18:209-219 |
| بيانات النشر: | American Chemical Society (ACS), 2016. |
| سنة النشر: | 2016 |
| مصطلحات موضوعية: | 0301 basic medicine, medicine.medical_specialty, Microscope, High-throughput screening, Protein Array Analysis, Nanotechnology, Bead, 01 natural sciences, law.invention, 03 medical and health sciences, Peptide Library, law, Drug Discovery, Fluorescence microscope, medicine, Combinatorial Chemistry Techniques, CMOS sensor, 010405 organic chemistry, Drug discovery, Chemistry, General Chemistry, General Medicine, Combinatorial chemistry, Microspheres, High-Throughput Screening Assays, 0104 chemical sciences, Spectral imaging, Autofluorescence, 030104 developmental biology, Microscopy, Fluorescence, visual_art, visual_art.visual_art_medium, Protein Binding |
| الوصف: | The increasing involvement of academic institutions and biotech companies in drug discovery calls for cost-effective methods to identify new bioactive molecules. Affinity-based on-bead screening of combinatorial one-bead one-compound libraries combines a split-mix synthesis design with a simple protein binding assay operating directly at the bead matrix. However, one bottleneck for academic scale on-bead screening is the unavailability of a cheap, automated, and robust screening platform that still provides a quantitative signal related to the amount of target protein binding to individual beads for hit bead ranking. Wide-field fluorescence microscopy has long been considered unsuitable due to significant broad spectrum autofluorescence of the library beads in conjunction with low detection sensitivity. Herein, we demonstrate how such a standard microscope equipped with LED-based excitation and a modern CMOS camera can be successfully used for selecting hit beads. We show that the autofluorescence issue can be overcome by an optical image subtraction approach that yields excellent signal-to-noise ratios for the detection of bead-associated target proteins. A polymer capillary attached to a semiautomated bead-picking device allows the operator to efficiently isolate individual hit beads in less than 20 s. The system can be used for ultrafast screening of200,000 bead-bound compounds in 1.5 h, thereby making high-throughput screening accessible to a wider group within the scientific community. |
| تدمد: | 2156-8944 2156-8952 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::b8011075241a54bd94cb8fb356162e32 https://doi.org/10.1021/acscombsci.5b00175 |
| رقم الأكسشن: | edsair.doi.dedup.....b8011075241a54bd94cb8fb356162e32 |
| قاعدة البيانات: | OpenAIRE |
Find this issue from the American Chemical Society | تدمد: | 21568944 21568952 |
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