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Using Raman spectroscopy to characterize biological materials

DOI: 10.1038/nprot.2016.036 DOI Help

Authors: Holly J. Butler (Lancaster University) , Lorna Ashton (Department of Chemistry, Lancaster University) , Benjamin Bird (Daylight Solutions) , Gianfelice Cinque (Diamond Light Source) , Kelly Curtis (Department of Biomedical Physics, Physics and Astronomy, University of Exeter) , Jennifer Dorney (Department of Biomedical Physics, Physics and Astronomy, University of Exeter) , Karen Esmonde-white (Department of Internal Medicine, University of Michigan Medical School) , Nigel J Fullwood (Department of Biomedical and Life Sciences, School of Health and Medicine, Lancaster University) , Benjamin Gardner (Department of Biomedical and Life Sciences, School of Health and Medicine, Lancaster University) , Pierre L Martin-hirsch (Lancaster Environment Centre, Lancaster University) , Michael J Walsh (University of Illinois at Chicago) , Martin R Mcainsh , Nicholas Stone (Gloucestershire Royal Hospital/Cranfield Uni) , Francis L. Martin (Lancaster Environment Centre, Lancaster University)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Protocols , VOL 11 , PAGES 664 - 687

State: Published (Approved)
Published: March 2016

Abstract: Raman spectroscopy can be used to measure the chemical composition of a sample, which can in turn be used to extract biological information. Many materials have characteristic Raman spectra, which means that Raman spectroscopy has proven to be an effective analytical approach in geology, semiconductor, materials and polymer science fields. The application of Raman spectroscopy and microscopy within biology is rapidly increasing because it can provide chemical and compositional information, but it does not typically suffer from interference from water molecules. Analysis does not conventionally require extensive sample preparation; biochemical and structural information can usually be obtained without labeling. In this protocol, we aim to standardize and bring together multiple experimental approaches from key leaders in the field for obtaining Raman spectra using a microspectrometer. As examples of the range of biological samples that can be analyzed, we provide instructions for acquiring Raman spectra, maps and images for fresh plant tissue, formalin-fixed and fresh frozen mammalian tissue, fixed cells and biofluids. We explore a robust approach for sample preparation, instrumentation, acquisition parameters and data processing. By using this approach, we expect that a typical Raman experiment can be performed by a nonspecialist user to generate high-quality data for biological materials analysis.

Subject Areas: Physics


Instruments: B22-Multimode InfraRed imaging And Microspectroscopy

Added On: 02/11/2016 11:47

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