The use of Raman spectroscopy combined with saliva is an exciting emerging spectroscopy-biofluid combination. In this review, we summarize current methods employed in such studies, in particular the collection, pretreatment, and storage of saliva, as well as measurement procedures and Raman parameters used. Given the need for sensitive detection, surface-enhanced Raman methods are also surveyed, alongside chemometric techniques. A meta-analysis of variables is compiled. We observe a wide range of approaches and conclude that standardization of methods and progress to more extensive validation Raman-saliva studies is necessary. Nevertheless, the studies show tremendous promise toward the improvement of speed, diagnostic accuracy, and portable device possibilities in applications such as healthcare, law enforcement, and forensics.

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Yeasts constitute an oft-neglected class of pathogens among which the resistance to first-line treatments, attributed in part to mutations in efflux pumps, is rapidly emerging. Their thick, chitinreinforced cell walls render cell lysis difficult, complicating their analysis and identification by methods routinely used for bacteria, including matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Liquid extraction surface analysis mass spectrometry (LESA-MS) has previously been applied to the analysis of intact proteins from Gram-positive and Gram-negative bacterial colonies sampled directly on solid nutrient media. To date, a similar analysis of yeast colonies has not proved possible. Here we demonstrate the rapid release of intact yeast proteins for LESA-MS by electroporation using a home-built high-voltage device designed to lyse cells grown in colonies on agar media. Detection and identification of previously inaccessible proteins from baker's yeast Saccharomyces cerevisiae, as well as two clinically relevant yeast species (Candida glabrata and Cryptococcus neoformans), is shown. The electroporation approach also has the potential to be translated to other mass spectrometric analysis techniques, including MALDI and various ambient ionization methods.

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