Two-photon excitation STED microscopy

Moneron, Gael; Hell, Stefan W.

doi:10.1364/oe.17.014567

Cited by 210 publications

(141 citation statements)

References 21 publications

(25 reference statements)

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“…For this reason, several groups have extended STED microscopy to two-photon excitation using different optical systems and modalities, allowing imaging deeper into e.g. brain tissue slices [15,16,17,18].…”

Section: Major Technological Progress Since 2007mentioning

confidence: 99%

STED microscopy—towards broadened use and scope of applications

Blom

Widengren

2014

Current Opinion in Chemical Biology

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PostprintThis is the accepted version of a paper published in Current opinion in chemical biology. This paper has been peer-reviewed but does not include the final publisher proof-corrections or journal pagination.Citation for the original published paper (version of record):Blom, H., Widengren, J. (2014) STED microscopy -towards broadened use and scope of applications. Current opinion in chemical biology AbstractStimulated Emission Depletion (STED) microscopy has been convincingly demonstrated for fundamental studies in cells, living tissue and organisms. Today, a major trend in the STED technique development is to make the instruments simpler and more user-friendly, without compromising performance. This has become possible by new low-cost, turnkey laser technology and by implementing specifically designed phase plates and birefringent segmented polarization elements. With these simpler and cheaper realizations of STED now becoming more broadly available, and with a parallel development of sample preparation and fluorophore reporter molecules ultimately setting the limit of the image quality, contrast and resolution, we can expect a significant increase in the use of STED, in science as well as for clinical and drug development purposes. Highlights-STED imaging of cells, living tissue and organisms has been convincingly demonstrated -The STED technology has in the last 6-7 years become far easier to realize -This largely relies on new laser technology and customized optical elements -New fluorescent markers and sample preparation procedures support the development -By increased accessibility, a strong increase in the use of STED can be expected

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Section: Major Technological Progress Since 2007mentioning

confidence: 99%

STED microscopy—towards broadened use and scope of applications

Blom

Widengren

2014

Current Opinion in Chemical Biology

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“…Nonresonant vaporization has been applied in higher intensity regimes to induce ionization as well as vaporization, but has been limited to small molecules to date [40,41]. Nonlinear processes have also been used for a variety of techniques including stimulated emission depletion microscopy [42] and 2-photon microscopy [43], yielding enhanced spatial resolution over the traditional one photon process.…”

Section: Laser Femtosecond Laser Vaporization Of Membrane-derived Promentioning

confidence: 99%

Analysis of Amphiphilic Lipids and Hydrophobic Proteins Using Nonresonant Femtosecond Laser Vaporization with Electrospray Post-Ionization

Brady

Judge

Levis

2011

J. Am. Soc. Mass Spectrom.

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Amphiphilic lipids and hydrophobic proteins are vaporized at atmospheric pressure using nonresonant 70 femtosecond (fs) laser pulses followed by electrospray post-ionization prior to being transferred into a time-of-flight mass spectrometer for mass analysis. Measurements of molecules on metal and transparent dielectric surfaces indicate that vaporization occurs through a nonthermal mechanism. The molecules analyzed include the lipids 1-monooleoyl-rac-glycerol, 1,2-dihexanoyl-sn-glycero-3-phosphocholine, 1,2-dimyristoyl-sn-glycero-3-phosphocholine, and the hydrophobic proteins gramicidin A, B, and C. Vaporization of lipids from blood and milk are also presented to demonstrate that lipids in complex systems can be transferred intact into the gas phase for mass analysis.

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“…For example, two-photon excitation microscopy, an established method that allows researchers to see further into tissues, can be combined with STED microscopy. Earlier this year, Hell's group used the technique on mammalian cell nuclei tagged with fluorescent nano particles, obtaining a resolution of less than 70 nm in the focal plane 1 .…”

Section: Technology Tweaksmentioning

confidence: 99%

Ever-increasing resolution

Rae¹

2009

Nature

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Overcoming the limitations of spatial and temporal resolution to image within a cell is no easy feat. Kelly Rae Chi examines the latest diffraction-busting technologies.For many years it was a source of frustration for biologists that the internal components of a cell were practically invisible to them. Researchers believed that the wavelength of light determined a fundamental limit to the resolution of optical microscopes.However, it now seems that the wavelength of light was not such a limiting factor after all. Super-resolution technology allows researchers to see details that are difficult or impossible to image with conventional light microscopes -at resolutions of 100 nanometres or better."There's a huge explosion of interest and progress, " says W. E. Moerner, a professor of chemistry and applied physics at Stanford University in California. "That makes it very exciting to watch and to participate in. "Although the theory behind super-resolution is advancing, and many different techniques are now validated, commercialization of these new microscopes is to some extent in its infancyso much so that many research teams still tackle super-resolution using their own lenses, lasers and algorithms. Upping commercializationThe past year or so has been particularly active in terms of commercialization. Throughout 2008, Applied Precision of Issaquah, Washington, tested prototypes of its DeltaVision OMX system. The final version is available this year and is already installed in 11 laboratories. It uses three-dimensional structured-illumination microscopy, or 3D-SIM, which illuminates a sample with a series of light patterns that look like bar codes. The low-resolution light patterns reflect off the fine structure of the sample to create moiré fringes. By applying bar codes in different orientations and processing the reflections using computer algorithms, the microscope generates a high-resolution image of the underlying structure.Carl Zeiss, headquartered in Oberkochen, Germany, is also using the SIM method in its system, ELYRA S.1, to be launched in January. And the company is launching ELYRA P.1, which is based on photo activated localization microscopy (PALM). This uses photo activatable proteins that are scattered throughout the sample. Taking multiple frames of the photoactivated dots, and combining them into a high-resolution image, the ELYRA P.1 produces a resolution of around 20 nm. Zeiss also offers a single system, the ELYRA PS.1, that combines laser scanning microscopy, PALM and SIM.Leica Microsystems in Wetzlar, Germany, is using a variation of stimulated emission depletion (STED) microscopy in its newest super-resolution system. The technique uses a laser beam to excite fluorescent dyes, or fluorophores, within the sample. As with a normal laser scanning microscope, the size of the excited spot determines the resolution of the microscope. In STED microscopy, to improve resolution and narrow the focus of the beam, an excitatory laser pulse is immediately followed by a ring-shaped depletion laser pulse ...

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Two-photon excitation STED microscopy

Cited by 210 publications

References 21 publications

STED microscopy—towards broadened use and scope of applications

STED microscopy—towards broadened use and scope of applications

Analysis of Amphiphilic Lipids and Hydrophobic Proteins Using Nonresonant Femtosecond Laser Vaporization with Electrospray Post-Ionization

Ever-increasing resolution

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