Sammanfattning

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.

Publicerad i

Nature Protocols
2015, Volym: 10, nummer: 2, sidor: 217-240
Utgivare: NATURE PUBLISHING GROUP

SLU författare

• Lundberg-Felten, Judith

  • Institutionen för skoglig genetik och växtfysiologi, Sveriges lantbruksuniversitet
    

• Hall, Hardy

  • Institutionen för skoglig genetik och växtfysiologi, Sveriges lantbruksuniversitet
    

UKÄ forskningsämne

Biokemi och molekylärbiologi


Publikationens identifierare

DOI: https://doi.org/10.1038/nprot.2015.008

Permanent länk till denna sida (URI)

https://res.slu.se/id/publ/76145