Glioblastoma is the most malignant and common high-grade brain tumor with a 14-month overall survival length. According to recent World Health Organization Central Nervous System tumor classification (2021), the diagnosis of glioblastoma requires extensive molecular genetic tests in addition to the traditional histopathological analysis of Formalin- Fixed Paraffin-Embedded (FFPE) tissues. Time-consuming and expensive molecular tests as well as the need for clinical neuropathology expertise are the challenges in the diagnosis of glioblastoma. Hence, an automated and rapid analytical detection technique for identifying brain tumors from healthy tissues is needed to aid pathologists in achieving an errorfree diagnosis of glioblastoma in clinics. Here, we report on our clinical test results of Raman spectroscopy and machine learning-based glioblastoma identification methodology for a cohort of 20 glioblastoma and 18 white matter tissue samples. We used Raman spectroscopy to distinguish FFPE glioblastoma and white matter tissues applying our previously reported protocols about optimized FFPE sample preparation and Raman measurement parameters. One may analyze the composition and identify the subtype of brain tumors using Raman spectroscopy since this technique yields detailed molecule-specific information from tissues. We measured and classified the Raman spectra of neoplastic and non-neoplastic tissue sections using machine learning classifiers including support vector machine and random forest with 86.6% and 83.3% accuracies, respectively. These proof-of-concept results demonstrate that this technique might be eventually used in the clinics to assist pathologists once validated with a larger and more diverse glioblastoma cohort and improved detection accuracies.
The real-time polymerase chain reaction (RT-PCR) analysis using nasal swab samples is the gold standard approach for COVID-19 diagnosis. However, due to the high false-negative rate at lower viral loads and complex test procedure, PCR is not suitable for fast mass screening. Therefore, the need for a highly sensitive and rapid detection system based on easily collected fluids such as saliva during the pandemic has emerged. In this study, we present a surface-enhanced Raman spectroscopy (SERS) metasurface optimized with genetic algorithm (GA) to detect SARS-CoV-2 directly using unprocessed saliva samples. During the GA optimization, the electromagnetic field profiles were used to calculate the field enhancement of each structure and the fitness values to determine the performance of the generated substrates. The obtained design was fabricated using electron beam lithography, and the simulation results were compared with the test results using methylene blue fluorescence dye. After the performance of the system was validated, the SERS substrate was tested with inactivated SARS-CoV-2 virus for virus detection, viral load analysis, cross-reactivity, and variant detection using machine learning models. After the inactivated virus tests are completed, with 36 PCR positive and 33 negative clinical samples, we were able to detect the SARS-CoV-2 positive samples from Raman spectra with 95.2% sensitivity and specificity.
Raman spectroscopy is a highly sensitive and specific technique for identifying tissue compositions. Raman-based characterization of normal and abnormal tissues is impeded due to the variability in routine tissue preparation techniques, fluorescent background, and molecular heterogeneity. Thus, sample preparation and Raman measurement conditions for tissue sections must be optimized. Here, we present an optimized Raman protocol and sample preparation method for brain tissue sections. This protocol allows the characterization of tissues and recognition of brain tumors by refining laser power, accumulation/exposure times, excitation wavelength, glass/CaF2 substrate, deparaffinization solvent, and the thickness of sections.
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