研究目的
To assess the ability of Raman spectroscopy to screen for histologically confirmed cases of Cervical Intraepithelial neoplasia (CIN) using biobanked liquid based cytology (LBC) samples.
研究成果
Raman spectroscopy can effectively discriminate disease-free cervical LBC samples from those with CIN3, particularly using samples stored at -25°C. Pooling fresh and biobanked samples does not affect performance, demonstrating that biobanks are a valuable resource for Raman-based screening studies.
研究不足
Samples stored at -80°C are not suitable for Raman spectroscopy due to cell lysis and debris, limiting the use of such biobanked samples. The study had a small sample size for some comparisons (e.g., 5 samples for biobanked vs. fresh), and further research is needed with different biobank specimens.
1：Experimental Design and Method Selection:
The study compared Raman spectroscopy for detecting CIN in fresh and biobanked LBC samples stored at different temperatures (-80°C and -25°C). It used partial least squares discriminant analysis (PLS-DA) for data analysis.
2：Sample Selection and Data Sources:
Samples were collected from patients with no disease (cytology negative and HPV negative) and disease (histologically confirmed CIN3 with HPV positivity). Biobanked samples were from the Scottish HPV Archive, and fresh samples were from Coombe Women and Infants University Hospital.
3：List of Experimental Equipment and Materials:
ThinPrep 2000 processor (Hologic Inc.), HORIBA Jobin Yvon XplorRA Raman system with Olympus microscope BX41, X100 objective, 532 nm diode laser, CCD detector, PreservCyt solution, glass slides, and Matlab software for data analysis.
4：Experimental Procedures and Operational Workflow:
Samples were prepared using the ThinPrep processor, Raman spectra were acquired from cell nuclei, data was pre-processed (smoothing, baseline correction, normalization), and PLS-DA was applied with leave-one-patient-out cross-validation.
5：Data Analysis Methods:
Data was analyzed using PLS-DA to discriminate between sample groups, with sensitivity and specificity calculated for CIN3 detection.
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