Optimization of measurement mode and sample processing for FTIR microspectroscopy in skin cancer research

Area of Science:

• Biomedical Optics
• Molecular Spectroscopy
• Cancer Research

Background:

• Fourier Transform Infrared (FTIR) microspectroscopy offers molecular-level insights into cells and tissues.
• Common FTIR microspectroscopy modes include transmission and transflection, often using paraffinized or deparaffinized samples.
• Variability in measurement modes and sample processing impacts spectral analysis, lacking a standardized protocol for optimal classification.

Purpose of the Study:

• To compare FTIR microspectroscopy of melanoma cell lines (IPC-298 and SK-MEL-30) using transmission and transflection modes.
• To evaluate the impact of paraffinized versus deparaffinized sample processing on spectral data.
• To determine the optimal combination of measurement mode and sample processing for accurate cell classification.

Main Methods:

• Acquisition of FTIR spectra from primary (IPC-298) and metastatic (SK-MEL-30) melanoma cell lines.
• Measurement in both transmission and transflection modes with both paraffinized and deparaffinized samples.
• Development of a Partial Least Squares Discriminant Analysis (PLS-DA) model for cell line classification.

Main Results:

• Significant spectral differences were observed based on measurement mode and deparaffinization status.
• The PLS-DA model achieved high classification accuracy across all tested conditions.
• Paraffinized samples analyzed in transflection mode demonstrated superior classification performance.

Conclusions:

• FTIR microspectroscopy, using various modes and sample preparations, can effectively differentiate melanoma cell lines.
• The biochemical signatures for discriminating cell lines are robust across different methods.
• Transflection mode with paraffinized samples is recommended for optimal classification accuracy in this context.