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Hierarchical imaging: a new concept for targeted imaging of large volumes from cells to tissues

Wacker, Irene; Spomer, Waldemar; Hofmann, Andreas; Thaler, Marlene; Hillmer, Stefan; Gengenbach, Ulrich; Schröder, Rasmus R.

Abstract (englisch):
Background: Imaging large volumes such as entire cells or small model organisms at nanoscale resolution seemed
an unrealistic, rather tedious task so far. Now, technical advances have lead to several electron microscopy (EM)
large volume imaging techniques. One is array tomography, where ribbons of ultrathin serial sections are deposited
on solid substrates like silicon wafers or glass coverslips.
Results: To ensure reliable retrieval of multiple ribbons from the boat of a diamond knife we introduce a substrate
holder with 7 axes of translation or rotation specifically designed for that purpose. With this device we are able to
deposit hundreds of sections in an ordered way in an area of 22 × 22 mm, the size of a coverslip. Imaging such
arrays in a standard wide field fluorescence microscope produces reconstructions with 200 nm lateral resolution
and 100 nm (the section thickness) resolution in z.
By hierarchical imaging cascades in the scanning electron microscope (SEM), using a new software platform, we
can address volumes from single cells to complete organs. In our first example, a cell population isolated from
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Zugehörige Institution(en) am KIT Institut für Angewandte Informatik (IAI)
Publikationstyp Zeitschriftenaufsatz
Jahr 2016
Sprache Englisch
Identifikator DOI: 10.1186/s12860-016-0122-8
ISSN: 1471-2121
KITopen ID: 1000065550
HGF-Programm 43.22.03; LK 01
Erschienen in BMC cell biology
Band 17
Heft 1
Seiten 17:38
Schlagworte Array tomography, Serial sectioning, Section libraries, Hierarchical imaging, Large volume 3D reconstruction
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