MicroCT protocols for scanning egg capsules of Hexaplex trunculus
Sep 03, 2021

MicroCT protocols for scanning egg capsules of Hexaplex trunculus

  • 1Hellenic Centre for Marine Research
DISCLAIMER – FOR INFORMATIONAL PURPOSES ONLY; USE AT YOUR OWN RISK

The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to protocols.io is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment. Any action you take or refrain from taking using or relying upon the information presented here is strictly at your own risk. You agree that neither the Company nor any of the authors, contributors, administrators, or anyone else associated with protocols.io, can be held responsible for your use of the information contained in or linked to this protocol or any of our Sites/Apps and Services.
Micro-computed tomography (micro-CT) is a high-resolution 3D-imaging technique which is now increasingly applied in biological studies focusing on taxonomy and functional morphology. The creation of virtual representations of specimens can increase availability of otherwise underexploited and inaccessible samples. This protocol aims to standardise micro-CT scanning procedures for the egg capsules of the marine gastropod species Hexaplex trunculus.
 
Eva Chatzinikolaou, Kleoniki Keklikoglou 2021. MicroCT protocols for scanning egg capsules of Hexaplex trunculus. protocols.iohttps://dx.doi.org/10.17504/protocols.io.bxw5ppg6
ECCO project, Hellenic Foundation for Research and Innovation ( HFRI)
Grant ID: project ID 343
microCT, gastropods, egg capsules, Hexaplex trunculus, scanning
 protocol  ,  
Sep 02, 2021
Sep 03, 2021
 
52925
DISCLAIMER – FOR INFORMATIONAL PURPOSES ONLY; USE AT YOUR OWN RISK

The protocol content here is for informational purposes only and does not constitute legal, medical, clinical, or safety advice, or otherwise; content added to protocols.io is not peer reviewed and may not have undergone a formal approval of any kind. Information presented in this protocol should not substitute for independent professional judgment, advice, diagnosis, or treatment. Any action you take or refrain from taking using or relying upon the information presented here is strictly at your own risk. You agree that neither the Company nor any of the authors, contributors, administrators, or anyone else associated with protocols.io, can be held responsible for your use of the information contained in or linked to this protocol or any of our Sites/Apps and Services.

Sample preparation

1w 5d 1h 4m
1
5d
Fixation of egg capsules of Hexaplex trunculus in 5% formaldehyde buffered with seawater.


2
1h
Wash egg capsules with distilled water and dehydrated them with ethanol in gradually increasing concentrations (20%, 50%, 70%, 96%).
3
1w
Staining of egg capsules with 1% iodine in 96% ethanol.
4
2m
Washing of stained samples with 96% ethanol.
5
2m
Placement of samples inside a plastic Falkon tube with 96% ethanol as a scanning medium.

microCT scanning

3h 38m
6
3h 38m

SkyScan 1172
micro-computed tomographer (microCT)
Bruker
10L01170
Detail detectability: <1 μm Low contrast resolution (10% MTF): 5 μm Pixel size at maximum magnification: <0.8 μm X-ray source: Sealed microfocus X-ray tube, air cooled, >10,000h lifetime; Spot size <5μm @ 4W, 20-100kV, 0-250 μA (10W max) X-ray detector (camera): 11 Megapixel (4000X2300) 12-bit digital CCD-camera with fibre optic coupling to scintillator Maximum object size: 50 mm in diameter using offset scan Radiation safety: < 1 µSv/h at any point on the instrument surface


6.1
3h 38m
Scanning parameters for embryos and juvenile Hexaplex trunculus

Voltage: 80 kV
Current :124 μA
Filter: aluminium
Pixel size: 13.79 μm
Camera binning: 2 × 2
Exposure time: 1435 ms
Rotation: 360°
Rotation step: 0.40º
Frame averaging: 3

Images reconstruction

30m
7
30m
Projection images were reconstructed into cross sections using the SkyScan’s NRecon software (Bruker, Kontich, Belgium).

7.1
2m
Upload projection images.
7.2
2m
Perform X-Y alignment.
7.3
26m
Reconstruction parameters

Smoothing: 2
Ring artifact correction: 20
Beam hardening correction: 59%
Attenuation coefficients: 0 - 0.064
Save as: 16-bit TIFF images