Pic2Spec: Generative Modeling Reconstructs Single Cell Raman Fingerprints from Brightfield Images

2026-07-08

AI Summary

Although single-cell Raman spectroscopy provides label-free molecular fingerprints, its high-throughput application is hindered by slow acquisition speeds and reliance on specialized instrumentation. This study proposes a generative model that, for the first time, reconstructs high-fidelity virtual Raman spectra directly from conventional brightfield microscopy images. By learning a shared latent biochemical representation between cellular morphology and vibrational spectra, the method enables inference of molecular information without dedicated hardware. The generated spectra achieve cosine similarities of up to 98% and Pearson correlation coefficients of approximately 95% compared to experimentally measured data across both mammalian and bacterial cells. The approach accurately discriminates genetic modification status and predicts GFP expression, outperforming traditional image-based analysis by 20%, thereby transforming standard microscopes into platforms capable of molecular-level characterization.

Abstract

Single-cell molecular characterization remains a bottleneck in scalable biological analysis because of labeling requirements, limited multiplexing, and reagents that perturb physiology. Raman spectroscopy addresses these limits by providing chemically specific, label-free vibrational fingerprints, but long acquisition times and specialized instruments restrict high-throughput use. Here, we overcome this barrier by showing that spectral fingerprints can be reconstructed from brightfield microscopy using generative modeling. We introduce Pic2Spec, a framework that learns a shared latent biochemical representation linking image morphology to vibrational spectral structure, enabling virtual Raman spectroscopy without hardware. We validate Pic2Spec across mammalian and bacterial cells, generating high-fidelity spectra that reproduce measured Raman fingerprints with 98% cosine similarity and Pearson correlations of ~95%, while preserving biochemical peaks and population distributions. Beyond spectral similarity, Pic2Spec provides molecular-level resolution in bacterial systems: generated spectra discriminate mutation-driven transgenic states and predict GFP expression with accuracy approaching true Raman measurements, outperforming conventional image analysis by 20%. These findings establish Pic2Spec as a first demonstration of chemically informative virtual molecular fingerprinting from brightfield images, complementing slow, hardware-intensive spectroscopy with computational inference. By redefining microscopy as an inference-enabled molecular profiling platform, Pic2Spec democratizes label-free biochemical phenotyping and overcomes the hardware and time constraints that have confined spectroscopy to specialized laboratories. This enables high-throughput molecular analysis for clinical diagnostics, screening, and monitoring at the scale and accessibility of standard microscopy.

Problem

• single-cell molecular characterization
• Raman spectroscopy
• high-throughput analysis
• label-free
• brightfield microscopy

Innovation

• generative modeling
• virtual Raman spectroscopy
• brightfield microscopy
• single-cell molecular fingerprinting
• label-free phenotyping