Insect MicroCT Model Construction
A comprehensive guide to creating high-resolution 3D models of insect specimens using MicroCT technology.
Workflow Steps
1
Staining Process
Sample Preparation
- Anesthetize the insect sample using carbon dioxide (CO₂)
- Transfer the insect into a 1.5 mL centrifuge tube containing 1 mL of 0.5% Triton X-100 PBS (0.5% PBST)
- Close the tube with the cap and gently tap the tube on the table to allow the insect to settle at the bottom
- Incubate the insect for 5 minutes
Fixation
- Use 1 mL of Bouin's solution to fix the insect sample
- Fix for 16-24 hours
Washing
- After fixation, wash the insect sample 3 times in 1 mL PBS
- Each wash should last 30 minutes
Staining
- After washing, stain the insect sample with standard Lugol's solution
- Place the sample on a shaker and incubate for 5 days
Final Washing
- After the staining process, wash the insect sample in PBS
- Store it in PBS for scanning and analysis
- The sample should be scanned within one week after preparation
2
Sample Loading Setup
Sample Placement
- After staining, place the insect sample into PBS for scanning
- Seal the tip of a suitable-sized pipette tip and fill it with PBS
- Place the sample into the tip
- Seal the upper part of the pipette tip using a sealing film
Pipette Tip Material
- The pipette tip is made of polypropylene
- Has a thin wall (approximately 200-300 µm) and relatively low X-ray absorption
- The conical design ensures that the sample is stable during storage
3
Scanning Process
Scanning Setup
- Use a Dell workstation with the installed SkyScan software (Micro CT, v1.2.0, Bruker, Germany)
- Control the SkyScan 1272 high-resolution 3D X-ray microtomography scanner
X-ray Settings
- Set the X-ray source voltage to 35 kV
- Set the current to 40 µA
Sensor and Resolution
- Use a 1632×1092 pixel sensor
- Set to the maximum resolution that can still capture the entire insect sample
Random Movement Algorithm
- Set the random movement algorithm parameter to 30
- This corrects for any potential image blur caused by sample wobbling during rotation
Frame Averaging
- Set the average frames to 6
- This improves the signal-to-noise ratio
4
Reconstruction Process
Reconstruction Software
- The reconstruction of the Micro CT data is carried out using the NRecon software (Micro CT, v1.2.0, Bruker, Germany)
Influencing Factors
- The quality of the reconstruction is influenced by the image data
- This includes brightness and contrast of the captured images
Reconstruction Parameters
- Use the default shift correction to compensate for sample movement during scanning
- This addresses thermal fluctuation or slow sample movement
- Set circular artifact correction and X-ray beam hardening compensation to the default values
Output Settings
- The output images will be in 16-bit depth
Additional Resources
- SkyScan Software Documentation
Official documentation for SkyScan software and hardware
- Bruker MicroCT Academy
Training resources and webinars on MicroCT techniques
- InsectBase MicroCT Collection
Browse our collection of insect MicroCT models