Connected Microscopy with ZEISS

Single images are snapshots of a complex three-dimensional situation reduced to a 2D image. To fully understand the sample, volume data is necessary. ZEISS supports a broad variety of techniques to enter this 3D world: One option is ZEISS Array Tomography (AT): Here, a resin-embedded sample is cut into ultrathin serial sections where, the sequence of the sections defines the z-information of a subsequently computationally reconstructed 3D data set and the thickness of the sections determines the z-resolution of this z-stack. The thickness of the sections is typically between 40-100 nm. To make life much easier, a tape collecting ultramicrotome (ATUMtome) can be used to produce and collect the sections automatically on a continuous tape. Likewise, data acquisition, i.e. the imaging of the resulting series of sections, is a highly automated process with ZEISS AT. The 2D images are then computed into a 3D model. The serial sections can be stored and used for further imaging experiments.
As an alternative to sequentially cutting a block tissue into serial sections and subsequently imaging the sections, resin-embedded cells or tissues may be imaged in 3D directly within the SEM chamber in a fully automated workflow: the surface of a specimen block is repetitively cut away with images of the exposed block surface taken after each sectioning event (block face imaging). ZEISS offers two solutions to cut the sample within the scanning electron microscope: either using an ultramicrotome inside the SEM chamber (3View) or a focused ion beam (FIB)-SEM that combines a FE-SEM with a focused ion beam (FIB) for milling. While FIB-SEM is the best choice for higher z-resolution down to 3-4 nm, 3View in combination with Focal Charge Compensation provides robust and fastest imaging of 3D volumes with z-resolutions down to 15 nm. Focal Charge Compensation expands the versatility and considerably increases data quality without prolonging acquisition times and enables easy imaging of even the most charge-prone samples.

The multimodal data is automatically relocated and overlaid, and then stored in well-organized projects with intuitive image labels. The resulting data can be overlaid and stored together in one place which makes it easy to gain insights into the whole experiment and removes the difficulty associated with multiple storage locations and instruments. As a result, you gain efficiency and effectiveness with intuitive data management, simplified workflows and limitless navigation.
The figure presents an example from the neurosciences. Brain sections are labelled with GFP and Draq5 were imaged with a widefield system and a structure of interest was identified (A). Zoom in of the region of interest (B). The sample was transferred to an LSM equipped with an Airyscan and after retrieval the structure of interest this region was imaged in 3D with higher resolution (C). (D) shows the final 3D reconstruction of the z-stack taken with the LSM.

For example, the latest lab X-ray microscopes have moved beyond the geometric limitations imposed by classical microCT design, incorporating optical elements to achieve resolution and contrast comparable to many synchrotron experiments, both on the micro and nanoscale. In addition, they have even adopted increasing numbers of imaging modalities. Phase contrast imaging has extended the application space beyond the traditional absorption contrast to enable visualization of low or similar-density material phases, and diffraction contrast tomography (LabDCT) now offers nondestructive 3D grain mapping of crystalline samples for the first time.
Also similarly to the synchrotron community, laboratory X-ray tomography systems have leveraged the nondestructive nature of the technique to foster increasing development of various types of in situ and 4D imaging experiments. 4D (3D + time) experiments uniquely enable characterizing the evolution of a 3D structure via repeated imaging of the same sample as a function of sequential processing or external stimuli such as mechanical load, electric potential, wet or corrosive environment, different atmospheric conditions, and so on. 

The Airyscan detector from ZEISS provides nearly two times higher resolution in X, Y and Z and makes superresolution imaging easier than ever before – for any sample type and with any fluorophore.
Unlike conventional confocal techniques that claim to improve resolution through a partially closed pinhole and deconvolution calculations, Airyscan achieves greater resolution by opening the pinhole and collecting more light. During this Lunch and Learn, hear how ZEISS Airyscan detector helps you push sensitivity beyond the limits of conventional confocal microscopy and see examples of how researchers are using Airyscan to improve their imaging.

From large grains to tiny, fault-producing inclusions, the microstructure and chemistry of metals impact applications from infrastructure through automotive, aerospace, and consumer goods. Modern microscopy continues to evolve, just as metallurgical requirements keep challenging metallographers and analytical scientists with an increasing focus on data extraction from a variety of sources across a complex range of metallurgical samples. Multi-scale microscopy – spanning optical, x-ray, and electron technologies – has redefined microscopy beyond “great imaging,” coming of age as a single solution to access an array of analytical information.