Semianarium: Jaśniejsze barwniki fluorescencyjne: Poprawa wydajności kwantowej karbofluorescein i karborodamin przez inkorporacje małych pierścieni.

W piątek 9 X 2015 zapraszamy  o godz. 17 (sala: 2.163 CeNT) na seminarium Maćka Majewskiego, które będzie dotyczyło jego badań prowadzonych podczas stażu w Janelia Research Campus. Abstrakt:

Maciej Majewski, Jonathan B. Grimm and Luke D. Lavis

Analogi ksantenowych barwników znajdują wiele zastosowań w biologii i obrazowaniu. Poprzednie badania przeprowadzone w laboratorium Luka Lavisa w Janelii skutkowały stworzeniem serii nowych analogów fluoresceiny i rodaminy z węglem lub krzemem zastępującym centralny atom tlenu.1,2 Otrzymane barwniki wykazują poprawione właściwości spektroskopowe. Przesunięcie batochromowe o 50 nm umożliwia zaawansowane eksperymenty obrazowania komórkowego, takie jak wielokolorowa super-rozdzielcza mikroskopia oraz śledzenie pojedynczych molekuł. Pomimo poprawy wielu parametrów, analogi wykazują obniżoną wydajność kwantową w porównaniu do wzorcowych barwników.

Podczas seminarium zaprezentuję wyniki 3-miesięcznego stażu w Janelia Research Campus działającym pod Instytutem Medycznym Howarda Hughesa. Naszą propozycją rozwiązania problemu wydajności kwantowej było wprowadzenie małych węglowych pierścieni w miejsce centralnego tlenu. Modyfikacja usztywnia strukturę i ogranicza ruchy, które mogą promować nieradiacyjne przejścia z wzbudzonego stanu elektronowego. Opracowana strategia zakładała syntezę analogów karbofluoresceiny z wprowadzonym cyklopropanem, cyklobutanem oraz cyklopentanem, a następnie przekształcenie ich w karborodaminy. Na koniec zbadaliśmy właściwości spektroskopowe, aby określić, jaki wpływ na wydajności kwantową barwników wywarła wprowadzona modyfikacja.

  1. Grimm, J.B., et al. „A general method to improve fluorophores for live-cell and single-molecule microscopy.” Nature methods 12.3 (2015): 244-250.
  2. Grimm, J.B., et al. „Carbofluoresceins and carborhodamines as scaffolds for high-contrast fluorogenic probes.” ACS chemical biology 8.6 (2013): 1303-1310.
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