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J Neurosci Methods. 2017 Apr 15;282:20-33. doi: 10.1016/j.jneumeth.2017.03.002. Epub 2017 Mar 4.

Automating cell detection and classification in human brain fluorescent microscopy images using dictionary learning and sparse coding.

Author information

1
Memory and Aging Center, University of California San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: maryana.alegro@ucsf.edu.
2
Memory and Aging Center, University of California San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: panos.theofilas@ucsf.edu.
3
Memory and Aging Center, University of California San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: austin.nguy@ucsf.edu.
4
Memory and Aging Center, University of California San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: alejandra.castcap@gmail.com.
5
Memory and Aging Center, University of California San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: bill.seeley@ucsf.edu.
6
Medical School of the University of São Paulo, Av. Reboucas 381, São Paulo, SP 05401-000, Brazil. Electronic address: heinsen@mail.uni-wuerzburg.de.
7
Computational Research Division, Lawrence Berkeley National Laboratory, 1 Cyclotron Rd, Berkeley, CA 94720, USA; Berkeley Institute for Data Science, University of California Berkeley, Berkeley, CA 94720, USA. Electronic address: dushizima@lbl.gov.
8
Memory and Aging Center, University of California San Francisco, 675 Nelson Rising Lane, San Francisco, CA 94158, USA. Electronic address: lea.grinberg@ucsf.edu.

Abstract

BACKGROUND:

Immunofluorescence (IF) plays a major role in quantifying protein expression in situ and understanding cell function. It is widely applied in assessing disease mechanisms and in drug discovery research. Automation of IF analysis can transform studies using experimental cell models. However, IF analysis of postmortem human tissue relies mostly on manual interaction, often subjected to low-throughput and prone to error, leading to low inter and intra-observer reproducibility. Human postmortem brain samples challenges neuroscientists because of the high level of autofluorescence caused by accumulation of lipofuscin pigment during aging, hindering systematic analyses. We propose a method for automating cell counting and classification in IF microscopy of human postmortem brains. Our algorithm speeds up the quantification task while improving reproducibility.

NEW METHOD:

Dictionary learning and sparse coding allow for constructing improved cell representations using IF images. These models are input for detection and segmentation methods. Classification occurs by means of color distances between cells and a learned set.

RESULTS:

Our method successfully detected and classified cells in 49 human brain images. We evaluated our results regarding true positive, false positive, false negative, precision, recall, false positive rate and F1 score metrics. We also measured user-experience and time saved compared to manual countings.

COMPARISON WITH EXISTING METHODS:

We compared our results to four open-access IF-based cell-counting tools available in the literature. Our method showed improved accuracy for all data samples.

CONCLUSION:

The proposed method satisfactorily detects and classifies cells from human postmortem brain IF images, with potential to be generalized for applications in other counting tasks.

KEYWORDS:

Dictionary learning; Image segmentation; Immunofluorescence; Microscopy; Postmortem human brain; Sparse models

PMID:
28267565
PMCID:
PMC5600818
DOI:
10.1016/j.jneumeth.2017.03.002
[Indexed for MEDLINE]
Free PMC Article

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