Near-infrared MINFLUX imaging enabled by suppression of fluorophore blinking DOI Creative Commons

Chinmaya Venugopal Srambickal,

Hanie Esmaeeli,

Joachim Piguet

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 28, 2024

Abstract MINimal photon FLUXes (MINFLUX) offers super-resolution microscopy (SRM) with nanometer localization precision, more relaxed fluorophore brightness and photostability requirements than for other SRM techniques. Nonetheless, low probabilities have been reported in several MINFLUX studies, a broader use of less bright photostable fluorophores, including near-infrared (NIR) fluorophores has difficult to realize. In this work, we identified blinking as main cause erroneous (and dismissed) localizations imaging devised strategies overcome these effects. We systematically studied the blinking/switching properties cyanine emitting far-red or NIR range, over typical time scales (µs-10ms), sample excitation conditions used imaging. By subsequent simulations representative procedures, found that trans-cis isomerization, particular photo-reduction can generate significant errors. However, errors could be suppressed by balanced redox buffers repetitive beam scans. Implementing strategies, replacing slower, intrinsic switching needed transient binding fluorophore-labelled DNA strands complementary attached targets (DNA-PAINT), first demonstrate NIR-MINFLUX precision. This work presents an overall strategy, where characterization make it possible design optimal conditions, opening imaging, well related studies.

Язык: Английский

Near-infrared MINFLUX imaging enabled by suppression of fluorophore blinking DOI Creative Commons

Chinmaya Venugopal Srambickal,

Hanie Esmaeeli,

Joachim Piguet

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 28, 2024

Abstract MINimal photon FLUXes (MINFLUX) offers super-resolution microscopy (SRM) with nanometer localization precision, more relaxed fluorophore brightness and photostability requirements than for other SRM techniques. Nonetheless, low probabilities have been reported in several MINFLUX studies, a broader use of less bright photostable fluorophores, including near-infrared (NIR) fluorophores has difficult to realize. In this work, we identified blinking as main cause erroneous (and dismissed) localizations imaging devised strategies overcome these effects. We systematically studied the blinking/switching properties cyanine emitting far-red or NIR range, over typical time scales (µs-10ms), sample excitation conditions used imaging. By subsequent simulations representative procedures, found that trans-cis isomerization, particular photo-reduction can generate significant errors. However, errors could be suppressed by balanced redox buffers repetitive beam scans. Implementing strategies, replacing slower, intrinsic switching needed transient binding fluorophore-labelled DNA strands complementary attached targets (DNA-PAINT), first demonstrate NIR-MINFLUX precision. This work presents an overall strategy, where characterization make it possible design optimal conditions, opening imaging, well related studies.

Язык: Английский

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