X-Ray Microscopy of the Larval Crustacean Brain

  • Jakob KriegerEmail author
  • Franziska Spitzner
Part of the Methods in Molecular Biology book series (MIMB, volume 2047)


Micro-computed X-ray tomography (μCT) coupled with visualization techniques such as three-dimensional reconstruction of internal morphological structures has opened up new pathways for analyzing the anatomy of nervous systems in intact specimens. The possibility for combining μCT with other techniques is one of the major advantages of μCT scanning, and the technical development of higher resolutions in lab-based μCT-scanners allows for investigating the anatomy of specimens in the sub-milimeter range. The European shore crab Carcinus maenas features a larval development over four zoeal and one megalopal stage with body lengths ranging from 500 μm to 2000 μm. The developing nervous system in the larvae of C. maenas is organized into a central brain which is connected via esophageal connectives with a ventral nerve chord and segmental ganglia. Since soft tissues such as the nervous tissues feature low contrasts compared to other tissues such as muscles or cuticularized body parts, the interpretation in μCT scans is challenging and needs some practice. The protocol described here is also applicable for larger specimens of a variety of species and spans over 2–3 days resulting in an image stack ready for postprocessing and visualization.


μCT Brain Contrast-enhancement 3D reconstruction Carcinus maenas Crustacean Larva 



We cordially thank Marie K. Hörnig for macrophotographs of specimen preparation as well as Steffen Harzsch for reviewing and constructive criticism of the first version of the manuscript. This work was supported by the German Science Foundation (Research Training Group 2010 RESPONSE, DFG INST 292/119-1 FUGG, and DFG INST 292/120-1 FUGG).


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© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  1. 1.Department of Cytology and Evolutionary Biology, Zoological Institute and MuseumUniversity of GreifswaldGreifswaldGermany

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