Normalization in MALDI-TOF Imaging Datasets of Proteins: Practical Considerations

Overview

Journal Anal Bioanal Chem

Specialty Chemistry

Date 2011 Apr 12

PMID 21479971

Citations 74

Authors

Soren-Oliver Deininger

Dale S Cornett

Rainer Paape

Michael Becker

Charles Pineau

Sandra Rauser

Axel Walch

Eryk Wolski

Affiliations

Soon will be listed here.

Abstract

Normalization is critically important for the proper interpretation of matrix-assisted laser desorption/ionization (MALDI) imaging datasets. The effects of the commonly used normalization techniques based on total ion count (TIC) or vector norm normalization are significant, and they are frequently beneficial. In certain cases, however, these normalization algorithms may produce misleading results and possibly lead to wrong conclusions, e.g. regarding to potential biomarker distributions. This is typical for tissues in which signals of prominent abundance are present in confined areas, such as insulin in the pancreas or β-amyloid peptides in the brain. In this work, we investigated whether normalization can be improved if dominant signals are excluded from the calculation. Because manual interaction with the data (e.g., defining the abundant signals) is not desired for routine analysis, we investigated two alternatives: normalization on the spectra noise level or on the median of signal intensities in the spectrum. Normalization on the median and the noise level was found to be significantly more robust against artifact generation compared to normalization on the TIC. Therefore, we propose to include these normalization methods in the standard "toolbox" of MALDI imaging for reliable results under conditions of automation.

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