研究目的
To differentiate between malignant and benign thyroid nodules by investigating collagen organization in their capsules using polarization-resolved second harmonic generation microscopy.
研究成果
PSHG microscopy combined with image processing can provide complementary information for differentiating between benign and malignant thyroid nodule capsules, potentially aiding in thyroid cancer diagnosis alongside traditional methods.
研究不足
The study is limited to a small number of samples (10 areas). The method requires specialized equipment and may not be readily available in all clinical settings. Further validation with larger datasets is needed.
1：Experimental Design and Method Selection:
Polarization-resolved second harmonic generation (PSHG) microscopy was used to image collagen organization. A theoretical curve was fitted to SHG intensity data to extract information on susceptibility tensor components and collagen fiber orientation.
2：Sample Selection and Data Sources:
Thyroid tissue sections from formalin-fixed, paraffin-embedded blocks were used, with H&E staining for identification and unstained sections for PSHG imaging. 10 areas were imaged: 3 thyroid capsules, 3 PTC capsules, and 4 FA capsules.
3：List of Experimental Equipment and Materials:
Modified Leica TCS SP confocal laser scanning microscope, mode-locked Ti:Sapphire laser (Tsunami, Spectra Physics), 40x 0.75 NA objective, photomultiplier, achromatic half-wave plate, filters (short-pass and band-pass), glass slides, formalin-fixed paraffin-embedded tissue sections.
4：75 NA objective, photomultiplier, achromatic half-wave plate, filters (short-pass and band-pass), glass slides, formalin-fixed paraffin-embedded tissue sections. Experimental Procedures and Operational Workflow:
4. Experimental Procedures and Operational Workflow: Tissue sections were prepared and imaged with PSHG at laser polarization angles from 0 to 180° in 20° steps. SHG signal was collected in the forward direction. A custom Matlab code was used for pixel-by-pixel fitting of SHG intensity to a theoretical model.
5：Data Analysis Methods:
Fitting quality was assessed using R2 coefficient. Images were computed for collagen orientation angle and susceptibility tensor element ratios. Histograms and statistical parameters (mode, mean, standard deviation) were analyzed.
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confocal laser scanning microscope
TCS SP
Leica
Used for polarization-resolved second harmonic generation microscopy imaging of tissue samples.
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Ti:Sapphire laser
Tsunami
Spectra Physics
Provided excitation for SHG microscopy with specific pulse characteristics.
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achromatic half-wave plate
Used to rotate the incident laser beam linear polarization.
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photomultiplier
Collected the SHG signal in the forward direction.
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condenser lens
Used to collect SHG signal with high numerical aperture.
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short-pass filter
Filtered the collected light to isolate SHG signal.
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band-pass filter
Filtered the collected light to isolate SHG signal.
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