A robust co-localisation measurement utilising z-stack image intensity similarities for biological studies.

PloS One
Yinhai WangPeter W Hamilton

Abstract

Co-localisation is a widely used measurement in immunohistochemical analysis to determine if fluorescently labelled biological entities, such as cells, proteins or molecules share a same location. However the measurement of co-localisation is challenging due to the complex nature of such fluorescent images, especially when multiple focal planes are captured. The current state-of-art co-localisation measurements of 3-dimensional (3D) image stacks are biased by noise and cross-overs from non-consecutive planes. In this study, we have developed Co-localisation Intensity Coefficients (CICs) and Co-localisation Binary Coefficients (CBCs), which uses rich z-stack data from neighbouring focal planes to identify similarities between image intensities of two and potentially more fluorescently-labelled biological entities. This was developed using z-stack images from murine organotypic slice cultures from central nervous system tissue, and two sets of pseudo-data. A large amount of non-specific cross-over situations are excluded using this method. This proposed method is also proven to be robust in recognising co-localisations even when images are polluted with a range of noises. The proposed CBCs and CICs produce robust co-localisation ...Continue Reading

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Citations

Jan 22, 2015·Cytometry. Part a : the Journal of the International Society for Analytical Cytology·Thibault LagacheJean-Christophe Olivo-Marin

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Methods Mentioned

BETA
confocal microscopy

Software Mentioned

LAS AF

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