Micro-Computed Tomography (micro-CT) of Insect Brain Protocol

Application Note for Leica EM CPD300 - Life Science Research

October 06, 2016

Introduction
Species: Blow fly (Lucilia caesar)
Critical point drying of the blow fly with subsequent X-ray micro-computed tomography (micro-CT) to detect neuroanatomical features.

Procedure
Sample Holder:
Samples were placed individually in the chambers of Arthropoda holder.



Fixation and Dehydration

Bouin's fixativeovernight
0.1 M phosphate buffer (1.8% Sucrose, pH 7.2)3x10 min.
Ethanol series: 60%, 70%, 80%, 90%, 96%, 100%2x10 min.
Iodine staining (1% iodine solution in 100% ethanol)overnight
Ethanol: 100%2x10 min.

Mounting and Scanning
Dried samples were glued on insect pins and scanned with an Xradia MicroXCT-200 X-ray imaging system (Carl Zeiss X-ray Microscopy Inc., Pleasanton, USA).

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Leica EM CPD300 auto Program:

Fig.1: Volume reconstructions and virtual sections of the head and brain of a blow fly.

Mounting and Scanning
Dried samples were glued on insect pins and scanned with an Xradia MicroXCT-200 X-ray imaging system (Carl Zeiss X-ray Microscopy Inc., Pleasanton, USA).

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