RRC ID 89769
著者 Sakamoto J, Ishii H, Otomo K, Kozawa Y, Sato S, Yokoyama H, Nemoto T.
タイトル Wavelength-tunable depletion expands fluorophore compatibility in two-photon STED nanoscopy.
ジャーナル Biomed Opt Express
Abstract We report the development of a novel two-photon excitation stimulated emission depletion microscopy system, namely 2PE-scSTED, featuring a custom light source setup that integrates a supercontinuum (SC) laser light source with our proprietary 1064 nm pulsed semiconductor excitation laser. This SC source provides the capability of emitting a broadband spectral range, enabling highly reliable selection of depletion wavelength. Utilizing 2PE-scSTED, we investigated the depletion characteristics of several fluorophores. By employing a tunable filter to precisely select depletion wavelength from 640 to 800 nm, we systematically measured the depletion efficiency and identified the optimal wavelengths that maximize fluorescence depletion efficiency for four distinct fluorescent probes. This approach yielded super-resolution images with a lateral spatial resolution as fine as 45 nm (approximately λ2PE /24) with a relatively lower depletion power than our previous reports, and can be principally applicable to all fluorescent probes. Our results demonstrate that the integration of a broadband SC source removes the spectral constraint of fluorescence depletion, enabling tailored 2PE-STED microscopy for individual fluorophores. This enhances the versatility and applicability of 2PE-STED microscopy for super-resolution imaging of biological specimens.
巻・号 17(8)
ページ 4131-4144
公開日 2026-8-1
DOI 10.1364/BOE.603553
PII boe-17-8-4131
PMID 42609927
PMC PMC13481070
オルトメトリクス指標
オルトメトリクス指標項目
最多言及媒体 X(Twitter)
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過去6か月間でのオルトメトリクス指標の変動値 19.5
リソース情報
ヒト・動物細胞 HeLa(RCB0007)