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Micro-CT of Insect Larva Protocol

Application Note for Leica EM CPD300 - Life Science Research

Introduction
Species: red blood worm (midge larva)
Critical point drying of midge larvae with subsequent X-ray micro-computed tomography (micro-CT) to reconst­ruct the inner anatomy.

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

Fixation and Dehydration

2.5 % Glutardialdehyde (in 0.1 M phosphate buffer)overnight
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.

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Leica EM CPD300 auto Program:

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

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Fig.1: Volume reconstructions and virtual section of the red blood worm showing a variety of organ systems.

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