Evaluating different deep learning models for efficient extraction of Raman signals from CARS spectra.

The nonresonant background (NRB) contribution to the coherent anti-Stokes Raman scattering (CARS) signal distorts the spectral line shapes and thus degrades the chemical information. Hence, finding an effective approach for removing NRB and extracting resonant vibrational signals is a challenging task. In this work, a bidirectional LSTM (Bi-LSTM) neural network is explored for the first time to remove the NRB in the CARS spectra automatically, and the results are compared with those of three DL models reported in the literature, namely, convolutional neural network (CNN), long short-term memory (LSTM) neural network, and very deep convolutional autoencoders (VECTOR). The results of the synthetic test data have shown that the Bi-LSTM model accurately extracts the spectral lines throughout the range. In contrast, the other three models' efficiency deteriorated while predicting the peaks on either end of the spectra, which resulted in a 60 times higher mean square error than that of the Bi-LSTM model. The Pearson correlation analysis demonstrated that Bi-LSTM model performance stands out from the rest, where 94% of the test spectra have correlation coefficients of more than 0.99. Finally, these four models were evaluated on four complex experimental CARS spectra, namely, protein, yeast, DMPC, and ADP, where the Bi-LSTM model has shown superior performance, followed by CNN, VECTOR, and LSTM. This comprehensive study provides a giant leap toward simplifying the analysis of complex CARS spectroscopy and microscopy.

Effect of non-resonant background on the extraction of Raman signals from CARS spectra using deep neural networks.

We report the retrieval of the Raman signal from coherent anti-Stokes Raman scattering (CARS) spectra using a convolutional neural network (CNN) model. Three different types of non-resonant backgrounds (NRBs) were explored to simulate the CARS spectra viz (1) product of two sigmoids following the original SpecNet model, (2) Single Sigmoid, and (3) fourth-order polynomial function. Later, 50 000 CARS spectra were separately synthesized using each NRB type to train the CNN model and, after training, we tested its performance on 300 simulated test spectra. The results have shown that imaginary part extraction capability is superior for the model trained with Polynomial NRB, and the extracted line shapes are in good agreement with the ground truth. Moreover, correlation analysis was carried out to compare the retrieved Raman signals to real ones, and a higher correlation coefficient was obtained for the model trained with the Polynomial NRB (on average, ∼0.95 for 300 test spectra), whereas it was ∼0.89 for the other NRBs. Finally, the predictive capability is evaluated on three complex experimental CARS spectra (DMPC, ADP, and yeast), where the Polynomial NRB model performance is found to stand out from the rest. This approach has a strong potential to simplify the analysis of complex CARS spectroscopy and can be helpful in real-time microscopy imaging applications.