Rapid nonlinear image scanning microscopy

Gregor, Ingo; Spiecker, Martin; Petrovsky, Roman; Großhans, Jörg; Ros, Robert; Enderlein, Joerg

doi:10.1038/nmeth.4467

Cited by 76 publications

(63 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In a similar manner to the procedure used by Gregor et al 26 Fourier reweighting for Q-ISM was done as follows:…”

Section: A Fourier Reweighting Protocol For Q-ism Imagesmentioning

confidence: 99%

“…Some variants offer the same resolution improvement within an alloptical setup reducing the need in computational power and fast imagers [22][23][24] . Although modest, this resolution improvement is robust to changes in the sample and label type and can be integrated with additional microscopy modalities 25,26 .We present here a new super-resolution scheme, quantum image scanning microscopy (Q-ISM), utilizing the measurement of quantum correlations in an ISM architecture. During a confocal scan, each pair of detectors in a detector array generates a sharp image using photon correlations.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Super-resolution enhancement by quantum image scanning microscopy

et al. 2018

View full text Add to dashboard Cite

The principles of quantum optics have yielded a plethora of ideas to surpass the classical limitations of sensitivity and resolution in optical microscopy. While some ideas have been applied in proof-of-principle experiments, imaging a biological sample has remained challenging mainly due to the inherently weak signal measured and the fragility of quantum states of light. In principle, however, these quantum protocols can add new information without sacrificing the classical information and can therefore enhance the capabilities of existing super-resolution techniques. Image scanning microscopy (ISM), a recent addition to the family of super-resolution methods, generates a robust resolution enhancement without sacrificing the signal level. Here we introduce quantum image scanning microscopy (Q-ISM): combining ISM with the measurement of quantum photon correlation allows increasing the resolution of ISM up to two-fold, four times beyond the diffraction limit. We introduce the Q-ISM principle and obtain super-resolved optical images of a biological sample stained with fluorescent quantum dots using photon antibunching, a quantum effect, as a resolution enhancing contrast mechanism. Main TextThe diffraction limit, as formulated by Abbe, sets the attainable resolution in far-field optical microscopy to about half of the visible wavelength 1 , hindering its applicability in life science studies at very small scales. Over the past two decades, several super-resolution methods have successfully overcome the diffraction limit, including emission depletion microscopy, localization microscopy and structured illumination microscopy 2-6 . The continuous and rapid improvement in detector technology has enabled two more recent developments in the field of super-resolution microscopy, which are the center of this work: quantum super-resolution microscopy and image scanning microscopy (ISM). As for the first, a surge of interest in super-resolution imaging based on quantum optics concepts 7-13 , inspired and facilitated by the progress in high temporal resolution imagers, resulted in a few successful proof-of-principle demonstrations 7,8,14 . The second, ISM, relies on a small array of fast detector and offers a two-fold enhancement of resolution 15,16 . Since ISM is compatible with a standard confocal microscope architecture it has already been integrated into commercial products.While all super-resolution modalities violate at least one of the basic assumptions of the Abbe theory, many rely on breaking more than one. For instance, stimulated emission depletion (STED) and saturated structured illumination microscopy (SSIM) breach both the assumption of a linear response of a fluorophore to the excitation light and that of a uniform illumination field 17,18 . In contrast, the few demonstrations of quantum super-resolution microscopy 7,8,14 relied solely on violating the implicit assumption, underlying Abbe's derivation, that light behaves as waves rather than particles. ISM, as well, depends on violating a single assumption, a u...

show abstract

“…In a similar manner to the procedure used by Gregor et al 26 Fourier reweighting for Q-ISM was done as follows:…”

Section: A Fourier Reweighting Protocol For Q-ism Imagesmentioning

confidence: 99%

mentioning

confidence: 99%

Super-resolution enhancement by quantum image scanning microscopy

et al. 2018

View full text Add to dashboard Cite

show abstract

“…S.5) order SOFISM. 6,9,10,14,15,16,17,18,19,21,22,23 and 24 present irregular shapes in the 4th order SOFISM and were therefore not used for the estimate of resolution enhancement.…”

Section: S3 Resolution Estimate For Sofismmentioning

confidence: 99%

“…Re-scan confocal microscopy (RCM) [13] and optical photon reassignment microscopy (OPRA) [14] provide all-optical realisations of ISM that alleviate the need for a fast detector array and multiple exposures. Through PSF engineering [15,16] and two photon excitation [17,18,19,20], ISM was extended to achieve a longer depth of field, resistance to scattering and spectral multiplexing. Alternatively, one can apply the concept of pixel reassignment for the benefit of different types of microscopy contrasts.…”

Section: Introductionmentioning

confidence: 99%

Super-resolution optical fluctuation image scanning microscopy (SOFISM)

Sroda

Makowski

Rossman

et al. 2019

Frontiers in Optics + Laser Science APS/DLS

View full text Add to dashboard Cite

Super-resolution optical microscopy is a rapidly evolving scientific field dedicated to imaging sub-wavelength sized objects, leaving its mark in multiple branches of biology and technology. While several super-resolution optical microscopy methods have become a common tool in life science imaging, new methods, supported by cutting-edge technology, continue to emerge. One rather recent addition to the super-resolution toolbox, image scanning microscopy (ISM), achieves an up to twofold lateral resolution enhancement in a robust and straightforward manner. To further enhance ISM's resolution in all three dimensions, we present and experimentally demonstrate here super-resolution optical fluctuation image scanning microscopy (SOFISM). Measuring the fluorescence fluctuation contrast in an ISM architecture, we obtain images with a x2.5 lateral resolution beyond the diffraction limit along with an enhanced axial resolution for a fixed cell sample labeled with commercially available quantum dots. The inherent temporal averaging of the ISM technique enables image acquisition of the fluctuation correlation contrast within millisecond scale pixel dwell times. SOFISM can therefore offer a robust path to achieve high resolution images within a slightly modified confocal microscope, using standard fluorescent labels and within reasonable acquisition times.

show abstract

“…Although in general this strategy compensates for eventual aberrations and misalignments of the optical system, in the context of STED-ISM it also accommodates implicitly the strong dependency of the shift-vectors on the STED intensity. Conversely, other all-optical ISM implementations -both single-spot [17,20,21] or eventually multi-spots [22,23] architectures -should rely on proper modelling or prior calibration of the STED microscope's effective PSF, as a function of the STED beam intensity and of the general experimental conditions. To investigate the concrete advantages of STED-ISM over STED microscopy, we explored the conditions for which one is usually concerned about the maximum amount of stimulating photons delivered to the sample: live cell imaging ( Fig.…”

mentioning

confidence: 99%

Synergic Combination of Stimulated Emission Depletion Microscopy with Image Scanning Microscopy to Reduce Light Dosage

Tortarolo

Castello

Koho

et al. 2019

Preprint

View full text Add to dashboard Cite

Stimulated emission depletion (STED) microscopy is one of the most influential nanoscopy techniques; by increasing the STED beam intensity, it theoretically improves the spatial resolution to any desired value. However, the higher is the dose of stimulating photons, the stronger are the photo-bleaching and photo-toxicity effects, which potentially compromise live-cell and long-term imaging. For this reason the scientific community is looking for strategies to reduce the STED beam intensity needed to achieve a target resolution. Here, we show how the combination of STED microscopy with image scanning microscopy (ISM) meets this request. In particular, we introduce a new STED-ISM architecture -based on our recent single-photon-avalanche-diode (SPAD) detector array -which allows covering the near-diffraction limit resolution range with reduced STED beam intensity. We demonstrate this ability both with simulated data and in live-cell experiments. Because of (i) the minimal changes in the optical architecture of the typical point-scanning STED microscope; (ii) the parameter-free, robust and real-time pixel-reassignment method to obtain the STED-ISM image; (iii) the compatibility with all the recent progresses in STED microscopy, we envisage a natural and rapid upgrade of any STED microscope to the proposed STED-ISM architecture. * Corresponding authors: giuseppe.vicidomini@iit.it.

show abstract

Rapid nonlinear image scanning microscopy

Cited by 76 publications

References 27 publications

Super-resolution enhancement by quantum image scanning microscopy

Super-resolution enhancement by quantum image scanning microscopy

Super-resolution optical fluctuation image scanning microscopy (SOFISM)

Synergic Combination of Stimulated Emission Depletion Microscopy with Image Scanning Microscopy to Reduce Light Dosage

Contact Info

Product

Resources

About