Long-distance fluorescence lifetime imaging using stimulated emission

Dellwig, Thilo; Lin, Po‐Yen; Kao, Fu Jen

doi:10.1117/1.jbo.17.1.011009

Cited by 11 publications

(5 citation statements)

References 38 publications

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“…The well-known non-linear optical transient absorption and gain processes include two-photon absorption (TPA) [4][5][6], excited-state absorption (ESA) [5], ground-state depletion (GSD) [7], and stimulation emission (SE) [8]. Among these nonlinear optical processes, SE exhibits the novelties of subdiffraction imaging [9], detection of 'dark' (nonfluorescent) fluorophores [10], fluorescence imaging at an extended working distance [11][12][13], and polarization-resolved (PR) lifetime measurements of fluorophore [14]. SE signal is usually detected via lockin technique, in which the incident or pump beam is modulated and the non-linear signal of interest is extracted by demodulating the probe beam.…”

Section: Introductionmentioning

confidence: 99%

High temporal resolution and polarization resolved fluorescence lifetime measurements through stimulated emission

Rehman¹,

Das

Chen³

et al. 2020

Methods Appl. Fluoresc.

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We have implemented polarization-resolved fluorescence lifetime measurement through stimulated emission based pump-probe technique, which promises much higher temporal resolution (∼4 ps) than conventional time-correlated single-photon counting (TCSPC). The depolarization of ATTO 647N fluorescent dye is resolved through anisotropy fluorescence lifetime measurements, with variable time delay introduced between the pump and the probe beams. Importantly, the polarization anisotropy measurement and the corresponding rotational correlation time characterization of the fluorescent dye are carried out at various temperatures. We have also demonstrated the need of high temporal resolution via hetero Förster energy transfer (Hetero-FRET) through the interaction between the gold nanorods (GNRs) and the fluorescent dye ATTO 647N. Notably, our results compare highly favorably with conventional TCSPC method, which is rather limited in temporal resolution, for the above characterization. Additionally, this technique is applicable even under ambient light while being very cost-effective and robust.

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Section: Introductionmentioning

confidence: 99%

High temporal resolution and polarization resolved fluorescence lifetime measurements through stimulated emission

Rehman¹,

Das

Chen³

et al. 2020

Methods Appl. Fluoresc.

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“…Due to its coherent nature, SE does not attenuate like spontaneous fluorescence signal, which emits in 4π direction. SE was also demonstrated for long working distance detection [16,17]. Additionally, the integrated electronic time delay control of pump and probe beams enables precise lifetime measurement [18,19].…”

Section: Introductionmentioning

confidence: 99%

Background free imaging in stimulated emission fluorescence microscopy

Das

Chen

Rehman

et al. 2019

J. Opt.

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Recently, the pump-probe technique has found highly valued developments and applications in optical microscopy to study both labeled (fluorescence) and non-labeled (vibrational) for observation of cells and tissues. In this work, we use the mechanism of stimulated emission (SE) with dual frequency modulation to remove the large background signal that is originated from spontaneous emission. We have applied a high performance digital signal processing based lock-in detection for stimulated emission fluorescence microscopy. Critically, the pump beam is modulated at f1 and the probe beam is at a different frequency, f2, with the demodulation carried out at the sum of the two frequencies, (f1 + f2). In this way, the DC background is effectively removed. Note that the background is mainly attributed to the spontaneous emission of the pump beam, with modulated frequency f1.

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“…Henceforth its intensity decreases rapidly and inversely with the square of the distance, which would require the use of high numerical aperture (NA) optics to collect effectively. This problem can be remedied by stimulated emission (SE) based fluorescence detection [4][5][6][7][8] that maintains the molecule specific contrast through immuno-labeling while converting the incoherent spontaneous fluorescence signal into a coherent stimulated one. The stimulated fluorescence signal can then be collected effectively without high NA optics and the working distance is greatly extended.…”

Section: Introductionmentioning

confidence: 99%

Optical coherence detection with stimulated emission: a feasibility study

Chung

Kuo

et al. 2015

SPIE Proceedings

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A stimulated emission (SE) based optical coherence tomography (OCT) setup has been established for feasibility study. The setup conducts coherent gating for depth resolution using fluorescence based stimulated emission, an unprecedented scheme. The resulting depth resolution of this interferometric OCT setup is approximately 66 m, determined by the coherence of the stimulation light source. Additionally, the SE signal can be used to obtain fluorescence lifetime.

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Long-distance fluorescence lifetime imaging using stimulated emission

Cited by 11 publications

References 38 publications

High temporal resolution and polarization resolved fluorescence lifetime measurements through stimulated emission

High temporal resolution and polarization resolved fluorescence lifetime measurements through stimulated emission

Background free imaging in stimulated emission fluorescence microscopy

Optical coherence detection with stimulated emission: a feasibility study

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