دورية أكاديمية
أعرض في EDS

A Fluorescent Real-Time Plaque Assay Enables Single-Cell Analysis of Virus-Induced Cytopathic Effect by Live-Cell Imaging.

التفاصيل البيبلوغرافية
العنوان: A Fluorescent Real-Time Plaque Assay Enables Single-Cell Analysis of Virus-Induced Cytopathic Effect by Live-Cell Imaging.
المؤلفون: Arias-Arias JL; Centro de Investigación en Enfermedades Tropicales (CIET), Facultad de Microbiología, Universidad de Costa Rica, San José 11501-2060, Costa Rica.; Dulbecco Lab Studio, Residencial Lisboa 2G, Alajuela 20102, Costa Rica., Corrales-Aguilar E; Centro de Investigación en Enfermedades Tropicales (CIET), Facultad de Microbiología, Universidad de Costa Rica, San José 11501-2060, Costa Rica., Mora-Rodríguez RA; Centro de Investigación en Enfermedades Tropicales (CIET), Facultad de Microbiología, Universidad de Costa Rica, San José 11501-2060, Costa Rica.
المصدر: Viruses [Viruses] 2021 Jun 22; Vol. 13 (7). Date of Electronic Publication: 2021 Jun 22.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: MDPI Country of Publication: Switzerland NLM ID: 101509722 Publication Model: Electronic Cited Medium: Internet ISSN: 1999-4915 (Electronic) Linking ISSN: 19994915 NLM ISO Abbreviation: Viruses Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Basel, Switzerland : MDPI
مواضيع طبية MeSH: Optical Imaging/*instrumentation , Optical Imaging/*methods , Single-Cell Analysis/*methods , Viral Plaque Assay/*methods, Automation, Laboratory ; Cell Line ; Cytopathogenic Effect, Viral ; Single-Cell Analysis/instrumentation ; Viral Plaque Assay/instrumentation
مستخلص: Conventional plaque assays rely on the use of overlays to restrict viral infection allowing the formation of distinct foci that grow in time as the replication cycle continues leading to countable plaques that are visualized with standard techniques such as crystal violet, neutral red, or immunolabeling. This classical approach takes several days until large enough plaques can be visualized and counted with some variation due to subjectivity in plaque recognition. Since plaques are clonal lesions produced by virus-induced cytopathic effect, we applied DNA fluorescent dyes with differential cell permeability to visualize them by live-cell imaging. We could observe different stages of that cytopathic effect corresponding to an early wave of cells with chromatin-condensation followed by a wave of dead cells with membrane permeabilization within plaques generated by different animal viruses. This approach enables an automated plaque identification using image analysis to increase single plaque resolution compared to crystal violet counterstaining and allows its application to plaque tracking and plaque reduction assays to test compounds for both antiviral and cytotoxic activities. This fluorescent real-time plaque assay sums to those next-generation technologies by combining this robust classical method with modern fluorescence microscopy and image analysis approaches for future applications in virology.
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فهرسة مساهمة: Keywords: DNA fluorescent dyes; animal viruses; antiviral screening; automated image analysis; herpes simplex; live-cell imaging; plaque assay; real-time; vesicular stomatitis; yellow fever
تواريخ الأحداث: Date Created: 20210702 Date Completed: 20211209 Latest Revision: 20211214
رمز التحديث: 20221213
مُعرف محوري في PubMed: PMC8310316
DOI: 10.3390/v13071193
PMID: 34206483
قاعدة البيانات: MEDLINE
الوصف
تدمد:1999-4915
DOI:10.3390/v13071193