STED microscopy with a supercontinuum laser source.

Optics express (2008-06-26)
Dominik Wildanger, Eva Rittweger, Lars Kastrup, Stefan W Hell
ABSTRACT

We report on a straightforward yet powerful implementation of stimulated emission depletion (STED) fluorescence microscopy providing subdiffraction resolution in the far-field. Utilizing the same super-continuum pulsed laser source both for excitation and STED, this implementation of STED microscopy avoids elaborate preparations of laser pulses and conveniently provides multicolor imaging. Operating at pulse repetition rates around 1 MHz, it also affords reduced photobleaching rates by allowing the fluorophore to relax from excitable metastable dark states involved in photodegradation. The imaging of dense nanoparticles and of the microtubular network of mammalian cells evidences a spatial resolution of 30-50 nm in the focal plane, i.e. by a factor of 8-9 beyond the diffraction barrier.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Atto 647N NHS ester, BioReagent, suitable for fluorescence, ≥85% (coupling to amines)
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Phalloidin–Atto 647N, BioReagent, suitable for fluorescence, ≥85% (HPCE)
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Atto 532 NHS ester, BioReagent, suitable for fluorescence, ≥80% (coupling to amines)
Supelco
Atto 565 NHS ester, BioReagent, suitable for fluorescence, ≥70.0% (coupling to amines)
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Atto 647N, BioReagent, suitable for fluorescence, ≥98.0% (HPCE)
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Anti-Rabbit-IgG - Atto 647N antibody produced in goat, 1 mg/mL IgG
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Atto 565, BioReagent, suitable for fluorescence, ≥75% (sum of isomers, HPCE/HPLC)
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Anti-Mouse-IgG - Atto 647N antibody produced in goat, contains 50% glycerol as stabilizer
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Atto 425 NHS ester, BioReagent, suitable for fluorescence
Supelco
Atto 647N maleimide, BioReagent, suitable for fluorescence, ≥90% (coupling to thiols)
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Atto 647 NHS ester, BioReagent, suitable for fluorescence, ≥80% (coupling to amines)
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Atto 633, BioReagent, suitable for fluorescence
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Atto 565-Biotin, BioReagent, suitable for fluorescence, ≥90% (HPCE)
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Atto 647N DOPE, suitable for fluorescence
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Atto 647N-Streptavidin, BioReagent, suitable for fluorescence
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Atto 647N Protein Labeling Kit
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Atto 425
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NTA - Atto 647 N, BioReagent, suitable for fluorescence, ≥80% (HPCE)
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Atto 532, BioReagent, suitable for fluorescence, ≥90% (HPCE)
SAFC
Atto 465 NHS ester, BioReagent, suitable for fluorescence, ≥80% (HPCE)
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Atto 590-Biotin, BioReagent, suitable for fluorescence, ≥90.0% (HPCE)
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Phalloidin–Atto 390, suitable for fluorescence, ≥95.0% (HPCE)
Supelco
Atto 633 NHS ester, BioReagent, suitable for fluorescence, ≥90.0% (coupling to amines)
Supelco
Atto 590 NHS ester, BioReagent, suitable for fluorescence, ≥60% (coupling to amines)
Sigma-Aldrich
Atto 425-Biotin, BioReagent, suitable for fluorescence, ≥95.0% (HPCE)
Sigma-Aldrich
Atto 647N-Biotin, BioReagent, ≥95% (HPCE), suitable for fluorescence
Sigma-Aldrich
Atto 565 maleimide, BioReagent, suitable for fluorescence
Sigma-Aldrich
Atto 425-Streptavidin, BioReagent, suitable for fluorescence, ≥95% (HPCE)
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Atto 647N azide
Sigma-Aldrich
Phalloidin–Atto 532, suitable for fluorescence, ≥95.0% (HPCE)

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