The group of Siegfried Musser from Texas AM University recently published an article in Nature Cell Biology where they used a MIRAO 52E deformable mirror to perform nuclear pore complex imaging using adaptive optics in super resolution. Super resolution microscopy techniques, such as PALM and STORM, open the possibility to visualize the smallest intracellular components, lying well beyond the diffraction limit of light and which are not otherwise accessible using conventional fluorescence microscopy methods. One of such small intracellular structures, is a nuclear pore complex (NPC). Embedded in the nucleus membrane NPCs are massive multiprotein complexes that act as passageways for the transport of molecules into and out of the nucleus. With a molecular mass of 125 MDa in vertebrates, the NPC is one of the largest and most complex protein structures of eukaryotic cells and yet is still smaller than diffraction limit.

Breaking the diffraction limit

While numerous 3D light microscopy methods have been developed over the last few decades, single-molecule astigmatism imaging provides the highest spatial localization precision in X, Y and Z, and its useful Z-range matches well to that necessary to monitor cargo trafficking through NPCs. Although the simplest approach to achieve astigmatism imaging is via a cylindrical lens, here researchers used MIRAO 52E deformable mirror both to correct sample-induced aberrations and to add a small amount of astigmatism for 3D imaging. This way they demonstrated exceptional-quality calibration curves which ensured the highest localization precision in Z.  

Even though here researchers decided to implement standalone adaptive optics components, the same results could be obtained using MICAO 3DSR – a plug & play adaptive optics system from Imagine Optic. This device is compatible with any inverted-frame microscope and our MICAO 3DSR offer automatically includes installation services and long-term support in order to create a hassle-free experience for our customers.

If you’re interested in finding out more about our line of Microscopy and Adaptive Optics solutions, you can reach us at sales@imagine-optic.com or through the contact form (red enveloppe on the side).

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If you’re interested in finding out more about our line of Wavefront Sensors and Deformable Mirrors or Adative Optics solutions for Microscopy, you can reach us at sales@imagine-optic.com or through the contact form (red enveloppe on the side).

The post Adaptive Optics Light-Sheet Microscopy for functional Neuroimaging appeared first on Imagine Optic.

In order to circumvent this limitation and to enable dual-view, high-resolution light-sheet imaging, a team of researchers from Max Delbrück Center for Molecular Medicine (Germany), NIH and HHMI Janelia Research Campus (USA) recently proposed the use of adaptive optics to get rid of system aberrations (full publication here). Thanks to a smart symmetrical design and division of optical paths, aberrations can be corrected in both imaging paths using a single deformable mirror and sensorless iterative algorithms.

For all adaptive optics setups based on sensorless algorithms, a key performance requirement is the linearity of the phase modulator. The proposed method benefits greatly from the almost perfect linearity of the MIRA0 52E electromagnetic deformable mirror, as well as from its high dynamic range and intrinsic achromaticity when compared to most other phase modulators. If a particular mirror shape has to be kept for long-term imaging experiments, a specific version of MIRAO is also available, providing a stable shape within a few nm for hours (see MIRAO 52es).

The post Adaptive optics enables high-resolution, dual-view light-sheet microscopy through a tilted coverglass appeared first on Imagine Optic.

Recent innovations are stretching these limits. A publication in Nature Communications by the group of Lukas Kapitein from Utrecht University reports successful application of a novel merit function based on a Fourier transform of raw images, which, therefore, does not depend on the intensity and location of detections in every frame. Their method, which is called REALM, allows direct use of the “blinking images” of the PALM/STORM technique and permits detection of aberrations on the fly. The authors need as little as about 300 frames to detect aberrations, apply the correction using a deformable mirror and then acquire the PALM/STORM sequence using a perfect PSF. By correcting aberrations, in particular in depth, the number of counts per frame is demonstrated to be increased by a factor of at least 4, due to the restoration of the PSF quality.

Lukas Kapitein’s group is one of the leading teams in the world studying the cytoskeleton of neurons. They use several innovative research methods to understand the mechanisms by which cells establish and maintain their precise shape and intracellular organization. Their REALM technique in particular was made possible thanks to our MicAO 3DSR adaptive optics add-on for super resolution PALM/STORM systems. The method opens the door to deep SMLM in tissue with unprecedented 3D resolution.  For even more information about the method, (re)watch the webinar organized by Imagine Optic and presented by Marijn Siemons, PhD student in Lukas’s group, (and including a live demo!).

If you’re interested in finding out more about our AO solutions for Microscopy, you can reach us at sales@imagine-optic.com or through the contact form (red enveloppe on the side).

The post REALM, aberration correction for PALM/STORM microscopy – webinar appeared first on Imagine Optic.

The post EPFL choses MICAO 3DSR for best axial localization in microscopy appeared first on Imagine Optic.

We recently communicated the latest advances and results about this method and other implementations of our adaptive optics technology in microscopy at Focus on Microscopy, March 28-31 2021: the 5 talks we contributed to detailed how adaptive optics can boost imaging performance in Light-Sheet and Single-Molecule Localization Microscopy. 

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If you’re interested in finding out more about our  AO solutions for Microscopy, you can reach us at sales@imagine-optic.com or through the contact form (red enveloppe on the side).

The post (Re)Watch Webinar for Microscopy #2 – Adaptive Optics in SMLM for deep tissue imaging appeared first on Imagine Optic.

If you’re interested in finding out more about our line of Wavefront Sensors and Optical Metrology Systems and adaptive optics you can reach us at sales@imagine-optic.com or through the contact form (red enveloppe on the side).

The post New generation phase sensing and imaging: the MaxPhase project appeared first on Imagine Optic.

If you’re interested in finding out more about our line of Wavefront Sensors and Deformable Mirrors or AO solutions for Microscopy, you can reach us at sales@imagine-optic.com or through the contact form (red enveloppe on the side).

The post Adaptive Optics for Lattice Light Sheet microscopy – BIPSA 2020 appeared first on Imagine Optic.