Fluorescence depletion mechanisms in super-resolving STED microscopy

Rittweger, Eva; Rankin, Brian R.; Westphal, Volker; Hell, Stefan W.

doi:10.1016/j.cplett.2007.06.017

Cited by 72 publications

(54 citation statements)

References 17 publications

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“…16 Both processes compete with favorable ultrafast nonradiative S n f S 1 (or T n f T 1 ) relaxation followed by spontaneous or stimulated emission. 13 With these conditions in mind, we explored the use of rylene dyes for STED microscopy. In view of their large extinction coefficient, brightness, and photostability (especially when applied in materials sciences 4,17 ), these dyes should, at face value, be ideal for STED.…”

mentioning

confidence: 99%

“…In contrast, stimulated emission is instant, reversible, and so fundamental that almost any fluorophore can be transiently turned off by a STED beam. 13 Thus, if long-term darkening is minimized, STED should provide nanoscale resolution on the ensemble level with nearly any fluorophore. Therefore, avoiding any process that competes with stimulated emission is prudent.…”

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confidence: 99%

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Spectroscopic Rationale for Efficient Stimulated-Emission Depletion Microscopy Fluorophores

et al. 2010

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We report a rationale for identifying superior dyes for stimulated-emission depletion (STED) microscopy. We compared the dyes pPDI and pTDI, which displayed excellent photostability in single-molecule spectroscopy. Surprisingly, their photostability and performance in STED microscopy differed significantly. While single pTDI molecules could be visualized with excellent resolution (35 nm), pPDI molecules bleached rapidly under similar conditions. Femtosecond transient absorption measurements proved that the overlap between the stimulated-emission band and the excited-state absorption band is the main reason for the observed difference. Thus, assessment of the excited-state absorption band provides a rational means of dye selection and determination of the optimal wavelength for STED.

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confidence: 99%

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confidence: 99%

Spectroscopic Rationale for Efficient Stimulated-Emission Depletion Microscopy Fluorophores

et al. 2010

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“…[8][9][10] Conclusions STED itself has proven to be a revolutionary technology for farfield, noninvasive nanoscale imaging of fluorescently labeled structures. [47][48][49]51 Concurrently the LIFPA is a promising technique for velocity measurement. 43,53 Applying STED to LIFPA could establish a new method for nanoscopic velocity measurement that can 'side-step' the classical optical diffraction limit.…”

Section: Resultsmentioning

confidence: 99%

A novel far-field nanoscopic velocimetry for nanofluidics

Kuang

Wang

2010

Lab Chip

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For the first time we have been able to measure the flow velocity profile for nanofluidics with a spatial resolution better than 70 nm. Due to the diffraction resolution barrier, traditional optical methods have so far failed in measuring the velocity profile in a nanocapillary or a closed nanochannel without an opened sidewall. A novel optical point measurement method is presented which applies stimulated emission depletion (STED) microscopy to laser induced fluorescence photobleaching anemometer (LIFPA) techniques to measure flow velocity. Herein we demonstrate this far-field nanoscopic velocimetry method by measuring the velocity profile in a nanocapillary with an inner diameter of 360 nm. The closest measuring point to the wall is about 35 nm. This method opens up a new class of functional measuring techniques for nanofluidics and for nanoscale flows from the wall.

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“…The radiative transition in NVN crystal has been shown to occur with a high quantum yield of 0.95 [18]. Knowing the fluorescence decay time and the quantum yield of the transition, the stimulated emission cross section can be derived from the emission spectrum measurement [19]: [20]. The luminescence inhibition curve presented in Fig.…”

Section: Stimulated Emission Cross Sectionmentioning

confidence: 99%

STED imaging of green fluorescent nanodiamonds containing nitrogen-vacancy-nitrogen centers

Laporte¹,

Psaltis²

2015

Biomed. Opt. Express

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Abstract:We report Stimulated Emission Depletion (STED) imaging of green fluorescent nanodiamonds containing Nitrogen-Vacancy-Nitrogen (NVN) centers with a resolution of 70 nm using a commercial microscope. Nanodiamonds have been demonstrated to have the potential to be excellent cellular biomarkers thanks to their low toxicity and nonbleaching fluorescence, and are especially appealing for superresolution imaging technique like STED microscopy. However, only red fluorescent nanodiamonds containing Nitrogen-Vacancy (NV) centers have been used with STED microscopy so far. The existence of only one color nonbleaching center limits the possible observations, for instance it complicates spatial correlation studies with STED. To provide a nonbleaching probe in a different color, we characterize here the optical properties of the NVN defect for STED imaging. We demonstrate STED imaging of the green fluorescent nanodiamonds containing NVN centers, opening the door for long term two-color STED observation. Furthermore we exemplify the use of green nanodiamonds as a second color nonbleaching STED biomarker by imaging 70 nm fluorescent crystals up taken into HeLa cells.

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Fluorescence depletion mechanisms in super-resolving STED microscopy

Cited by 72 publications

References 17 publications

Spectroscopic Rationale for Efficient Stimulated-Emission Depletion Microscopy Fluorophores

Spectroscopic Rationale for Efficient Stimulated-Emission Depletion Microscopy Fluorophores

A novel far-field nanoscopic velocimetry for nanofluidics

STED imaging of green fluorescent nanodiamonds containing nitrogen-vacancy-nitrogen centers

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