María Gabriela Fernández-Manteca

Archaeological mollusk shells, such as those of Littorina obtusata/fabalis, hold valuable information about past human behavior and cultural practices. However, the original coloration of these shells, crucial for understanding their symbolic significance, is often lost due to taphonomic processes. Raman spectroscopy is a powerful technique for non-destructive analysis of archaeological samples, enabling the identification of pigments and mineralogical components. In this study, we present a methodology to predict, using Raman spectroscopy and k-means clustering, the original coloration of archaeological L. obtusata/fabalis shells which have lost their original coloration. Raman spectra were acquired from both modern shells, exhibiting a range of natural colors, and archaeological shell samples from La Chora cave (Cantabria, northern Spain). Spectral data were preprocessed to remove noise and baseline effects, and k-means clustering was applied to group the spectra based on their inherent spectral similarities. By comparing the spectral signatures of the archaeological samples with those of the modern shells within the generated clusters, we inferred the likely original coloration of the archaeological specimens. This approach provides a quantitative framework for predicting archaeological shell colors.

Spectroscopic techniques such as Surface-Enhanced Raman Scattering (SERS), Surface-Enhanced Infrared Absorption (SEIRA), and Surface-Enhanced Fluorescence (SEF) are essential analytical techniques used to study the composition of materials by analyzing the way materials scatter light, absorb infrared radiation or emit fluorescence signals. This provides information about their molecular structure and properties. However, traditional SERS, SEIRA, and SEF techniques can be limited in sensitivity, resolution, and reproducibility, hindering their ability to detect and analyze trace amounts of substances or complex molecular structures. Metasurfaces, a class of engineered two-dimensional metamaterials with unique optical properties, have emerged as a promising tool to overcome these limitations and enhance spectroscopic techniques. This article provides a state-of-the-art overview of metasurfaces for enhanced SERS, SEIRA and SEF, covering their theoretical background, different types, advantages, disadvantages, and potential applications.

Patella vulgata shells preserve geochemical and structural variations that can provide insights into past environmental conditions. Their composition, primarily calcium carbonate with organic residues from the biomineralization process, is influenced by external factors, such as sea surface temperature. Raman spectroscopy has emerged as a rapid, non-destructive tool for studying biogenic carbonates, enabling the identification of crystalline phases, organic components, and ion distribution. In this study, Raman imaging was applied to six shell sections of P. vulgata live-collected from Langre Beach in Cantabria, Spain. Spectral data were acquired using a Raman probe with a 532 nm excitation laser, providing high-resolution mapping of structural and compositional features. The analysis revealed spatial variations in mineralogy, organic matrix distribution, and ion incorporation in the calcium carbonate lattice, suggesting patterns originating during shell formation. Notably, the results suggest a consistent relationship between the organic and mineral components of the shells, with carotenoid distribution and carbonate ion substitution in the calcium carbonate lattice following similar growth patterns. These findings highlight the potential of Raman spectroscopy for studying biomineralization processes and the environmental records preserved in marine mollusk shells.

Water testing is becoming increasingly important due to dangerous phenomena such as Harmful Algal Blooms (HABs). Commonly, the content of a water sample is measured for the detection, monitoring and control of these events. Raman spectroscopy is a technique for the molecular characterization of materials in solid, liquid or gaseous form, which makes it an attractive method for analysing materials’ components. However, Raman scattering is a weak optical process and requires an accurate system for detection. In our work, we present, from design to fabrication, a microfluidic device on fused silica adapted to optimise the Raman spectrum of liquid samples when using a Raman probe. The device features a portable design for rapid on-site continuous flow measurements avoiding the use of large, costly and complex laboratory equipment. The main manufacturing technique used was ultrafast laser-assisted etching (ULAE). Finally, the effectiveness of the microfluidic device was demonstrated by comparing the Raman spectra of a known species of cyanobacteria with those obtained using other conventional substrates in laboratory analysis. The results demonstrate that the microfluidic device, under continuous flow conditions, exhibited a lower standard deviation of the Raman signal, reduced background noise and avoided signal variations caused by sample drying in static measurements.

Phytoplankton are essential to aquatic ecosystems but can cause harmful algal blooms (HABs) that threaten water quality, aquatic life, and human health. Developing new devices based on spectroscopic techniques offers a promising alternative for rapid and accurate monitoring of aquatic environments. However, phytoplankton undergo various physiological changes throughout their life cycle, leading to alterations in their optical properties, such as autofluorescence. In this study, we present a modification of a low-cost photobioreactor designed to implement fluorescence spectroscopy to analyze the evolution of spectral signals during phytoplankton growth cycles. This device primarily facilitates the characterization of changes in autofluorescence, providing valuable information for the development of future spectroscopic techniques for detecting and monitoring phytoplankton. Additionally, real-time testing was performed on cyanobacterial cultures, where changes in autofluorescence were observed under different conditions. This work demonstrates a cost-effective implementation of spectroscopic techniques within a photobioreactor, offering a preliminary analysis for the future development of functional field devices for monitoring aquatic ecosystems.