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
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
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.
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, focussed ion beam SEM or TEM can be performed. In particular, the Atlas 5 software ensures this process is streamlined in combination with ZEISS Crossbeam.
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
Y. Zhang et al. (2022), https://doi.org/10.3389/fcell.2022.880696
J. Ng. et al. (2016), https://doi.org/10.1038/srep38863