2D slice from a 3D XRM dataset, prepared for serial block-face SEM. Courtesy of Alana Burrell @EM_STP, CRICK Institute, United Kingdom

X-Ray Imaging Applications for Life Sciences

Streamline Your Multimodal Imaging Workflows

Screen Specimens for Quality and Identify Structures for Further Investigation

Generating high resolution, optimal datasets at the synchrotron or electron microscope requires perfectly prepared specimens, including staining and embedding. Streamlining acquisition routines depends on the selection of these optimal specimens and on identification of the regions of interest for higher resolution acquisition. Providing quick, non-destructive visualization of internal structure, high-resolution X-ray microscopy is ideal for both quality assessment and localization of regions of interest.

_{Courtesy of Alana Burrell @EM_STP, CRICK Institute, London}

Screen Specimens for Quality

2D slices from 3D reconstructions of two mouse olfactory bulb slices prepared for volume EM and imaged with the Versa XRM. Different sample preparation artefacts can be identified by screening to allow selection of the best specimens for subsequent analysis. Courtesy of Yuxin Zhang, the Francis Crick Institute, UK. For more information and examples please refer to ¹.

Efficient Verification of Specimen Quality Prior to Imaging at the Synchrotron or Electron Microscope

Variables in sample preparation, including fixing, staining, embedding and mounting, can all have negative impacts on sample quality and resulting imaging data¹. If problems with sample quality are only identified during the final steps of electron microscope or synchrotron acquisition, many hours and equipment rental fees have already been wasted capturing data that is ultimately unusable.

The ideal solution is to identify problems with specimen quality prior to the synchrotron or electron microscope (EM) so only optimal specimens are used for higher resolution imaging. This is where X-ray imaging is invaluable. Rapidly and non-destructively screening specimens in 3D and with high contrast to identify small imperfections or problems with staining ensures selection of the ideal specimens for subsequent analysis at higher resolution¹.

Precisely Target Your Acquisition Location

Bear jaw (120 mm x 200 mm) imaged from full jaw to micron-scale view of jaw-tooth interface. Macroscopically imaged using µCT with the ZEISS Flat Panel Detector to locate the interface of interest and the subsequent high-resolution acquisition with 0.4X and 4X objectives.

Generate a Multiscale 3D Map of Your Specimen to Identify Structures of Interest

With optimally prepared specimens selected, the next challenge is locating the precise region of interest for visualization at higher resolution. This task can be daunting as you are working with a relatively small field of view within a larger specimen. In addition, contrast enhancement often renders specimens opaque.

Non-destructive X-ray imaging is an easy way to generate a large, 3D specimen map which can be used to both explore internal structure and to guide your selection of a location for subsequent higher resolution acquisitions. Capturing information at multiple resolutions is straightforward with X-ray microscopy; you simply change the objective and zoom in with a higher magnification lens.

Correlated datasets of Drosophila brain, first imaged non-destructively using ZEISS Xradia Versa to identify the neurons of interest, and subsequently imaged using ZEISS Crossbeam to generate a high-resolution 3D volume of the neurons of interest. Courtesy of J. Ng, University of Cambridge, United Kingdom.²

Use the 3D High Resolution Map for Sample Trimming and Directed Acquisition

As you step to even higher resolution technologies, such as nano X-ray tomography or volume electron microscopy (vEM), these structural, overview maps are invaluable for directing the necessary sample trimming and precise location selection to start acquisition of the ultrastructural volume. And the same staining approaches used for vEM also work very well when imaging with X-rays, so no additional sample preparation is required.

The ZEISS Versa X-ray microscope provides a straightforward means of generating such sample maps for multimodal studies. From the high-resolution X-ray dataset, targeted acquisition of the region of interest using technologies such as serial blockface SEM, focused ion beam SEM or TEM can be performed. In particular, the Atlas 5 software ensures this process is streamlined in combination with ZEISS Crossbeam.

Courtesy of N. Senabulya and S. Smith, University of Michigan, USA.
Complete Cyclanthus bipartitus plant seed imaged with ZEISS Xradia Versa to locate the region for higher resolution acquisition (left). A zoom region cross-section of the ZEISS Xradia Versa data is shown (top right) with the same field of view captured at higher resolution using ZEISS Xradia Ultra (bottom right).

Complete Cyclanthus bipartitus plant seed Courtesy of N. Senabulya and S. Smith, University of Michigan, USA. — Courtesy of N. Senabulya and S. Smith, University of Michigan, USA.
Complete Cyclanthus bipartitus plant seed imaged with ZEISS Xradia Versa to locate the region for higher resolution acquisition (left). A zoom region cross-section of the ZEISS Xradia Versa data is shown (top right) with the same field of view captured at higher resolution using ZEISS Xradia Ultra (bottom right).

Extend your Multimodal X-ray Acquisition to the Nanoscale

X-ray imaging across length scale offers powerful insights into specimen structure. Providing structural information with spatial resolution down to 50 nm, ZEISS Xradia Ultra creates nanoscale 3D datasets. Generating the location map for the structures of interest using ZEISS Xradia Versa streamlines this multimodal imaging approach to achieve maximum efficiency and a hierarchical understanding of each specimen.

  
      Imaging in Action
      
    Francis Crick Institute, London

Learn how ZEISS Xradia Versa X-ray microscopes are used to significantly increase the efficiency of multimodal workflows that link neural function from in vivo light microscopy with ultrastructure that is captured at the synchrotron or using electron microscopy.

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