Your browser doesn't support javascript.
loading
Super-long single-molecule tracking reveals dynamic-anchorage-induced integrin function.
Tsunoyama, Taka A; Watanabe, Yusuke; Goto, Junri; Naito, Kazuma; Kasai, Rinshi S; Suzuki, Kenichi G N; Fujiwara, Takahiro K; Kusumi, Akihiro.
Afiliación
  • Tsunoyama TA; Membrane Cooperativity Unit, Okinawa Institute of Science and Technology Graduate University (OIST), Onna, Japan.
  • Watanabe Y; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, Japan.
  • Goto J; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, Japan.
  • Naito K; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, Japan.
  • Kasai RS; Institute for Frontier Life and Medical Sciences, Kyoto University, Kyoto, Japan.
  • Suzuki KGN; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, Japan.
  • Fujiwara TK; Center for Highly Advanced Integration of Nano and Life Sciences (G-CHAIN), Gifu University, Gifu, Japan.
  • Kusumi A; Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, Kyoto, Japan.
Nat Chem Biol ; 14(5): 497-506, 2018 05.
Article en En | MEDLINE | ID: mdl-29610485
Single-fluorescent-molecule imaging tracking (SMT) is becoming an important tool to study living cells. However, photobleaching and photoblinking (hereafter referred to as photobleaching/photoblinking) of the probe molecules strongly hamper SMT studies of living cells, making it difficult to observe in vivo molecular events and to evaluate their lifetimes (e.g., off rates). The methods used to suppress photobleaching/photoblinking in vitro are difficult to apply to living cells because of their toxicities. Here using 13 organic fluorophores we found that, by combining low concentrations of dissolved oxygen with a reducing-plus-oxidizing system, photobleaching/photoblinking could be strongly suppressed with only minor effects on cells, which enabled SMT for as long as 12,000 frames (~7 min at video rate, as compared to the general 10-s-order durations) with ~22-nm single-molecule localization precisions. SMT of integrins revealed that they underwent temporary (<80-s) immobilizations within the focal adhesion region, which were responsible for the mechanical linkage of the actin cytoskeleton to the extracellular matrix.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Integrinas / Colorantes Fluorescentes / Microscopía Fluorescente Límite: Animals / Humans Idioma: En Revista: Nat Chem Biol Asunto de la revista: BIOLOGIA / QUIMICA Año: 2018 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Integrinas / Colorantes Fluorescentes / Microscopía Fluorescente Límite: Animals / Humans Idioma: En Revista: Nat Chem Biol Asunto de la revista: BIOLOGIA / QUIMICA Año: 2018 Tipo del documento: Article País de afiliación: Japón Pais de publicación: Estados Unidos