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Vibrational spectroscopic image analysis of biological material using multivariate curve resolution–alternating least squares (MCR-ALS)
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Vibrational spectroscopic image analysis of biological material using multivariate curve resolution–alternating least squares (MCR-ALS)
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Journal Article
Vibrational spectroscopic image analysis of biological material using multivariate curve resolution–alternating least squares (MCR-ALS)
2015
Overview
Chemical compositional information can be extracted from Raman and infrared microscopic images by MCR-ALS. The algorithm finds the spectral profiles of compounds contributing to each image pixel and their relative concentrations.
Raman and Fourier transform IR (FTIR) microspectroscopic images of biological material (tissue sections) contain detailed information about their chemical composition. The challenge lies in identifying changes in chemical composition, as well as locating and assigning these changes to different conditions (pathology, anatomy, environmental or genetic factors). Multivariate data analysis techniques are ideal for decrypting such information from the data. This protocol provides a user-friendly pipeline and graphical user interface (GUI) for data pre-processing and unmixing of pixel spectra into their contributing pure components by multivariate curve resolution–alternating least squares (MCR-ALS) analysis. The analysis considers the full spectral profile in order to identify the chemical compounds and to visualize their distribution across the sample to categorize chemically distinct areas. Results are rapidly achieved (usually <30–60 min per image), and they are easy to interpret and evaluate both in terms of chemistry and biology, making the method generally more powerful than principal component analysis (PCA) or heat maps of single-band intensities. In addition, chemical and biological evaluation of the results by means of reference matching and segmentation maps (based on
k
-means clustering) is possible.
Publisher
Nature Publishing Group UK,Nature Publishing Group
Subject
/ Animals
/ Biochemistry and Molecular Biology
/ Biokemi
/ Biokemi och molekylärbiologi
/ Computational Biology/Bioinformatics
/ Image Processing, Computer-Assisted - methods
/ Islets of Langerhans - chemistry
/ Kemi
/ Methods
/ Pixels
/ Principal components analysis
/ protocol
/ Spectra
