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Hour-Long, Kilohertz Sampling Rate Three-Dimensional Single-Virus Tracking in Live Cells Enabled by StayGold Fluorescent Protein Fusions.
Lin, Yuxin; Exell, Jack; Lin, Haoting; Zhang, Chen; Welsher, Kevin D.
Afiliação
  • Lin Y; Department of Chemistry, Duke University, Durham, North Carolina 27708, United States.
  • Exell J; Department of Chemistry, Duke University, Durham, North Carolina 27708, United States.
  • Lin H; Department of Chemistry, Duke University, Durham, North Carolina 27708, United States.
  • Zhang C; Department of Chemistry, Duke University, Durham, North Carolina 27708, United States.
  • Welsher KD; Department of Chemistry, Duke University, Durham, North Carolina 27708, United States.
J Phys Chem B ; 128(23): 5590-5600, 2024 Jun 13.
Article em En | MEDLINE | ID: mdl-38808440
ABSTRACT
A viral infection process covers a large range of spatiotemporal scales. Tracking the viral infection process with fluorescent labels over long durations while maintaining a fast sampling rate requires bright and highly photostable labels. StayGold is a recently identified green fluorescent protein that has a greater photostability and higher signal intensity under identical illumination conditions compared to existing fluorescence protein variants. Here, StayGold protein fusions were used to generate virus-like particles (StayGold-VLPs) to achieve hour-long 3D single-virus tracking (SVT) with 1000 localizations per second (kHz sampling rate) in live cells. The expanded photon budget from StayGold protein fusions prolonged the tracking duration, facilitating a comprehensive study of viral trafficking dynamics with high temporal resolution over long time scales. The development of StayGold-VLPs presents a simple and general VLP labeling strategy for better performance in SVT, enabling exponentially more information to be collected from single trajectories and allowing for the future possibility of observing the entire life cycle of a single virus.
Assuntos

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Viroses / Proteínas de Fluorescência Verde Limite: Humans Idioma: En Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Viroses / Proteínas de Fluorescência Verde Limite: Humans Idioma: En Ano de publicação: 2024 Tipo de documento: Article