ArticleBiophysical journal2006
In situ background estimation in quantitative fluorescence imaging.
Article in Biophysical journal, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 13 papers.
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Who cites it
13 citing papers in PubMed.
- Automated detection of c-Fos-expressing neurons using inhomogeneous background subtraction in fluorescent images.Neurobiology of learning and memory · 2025Article
- Impact of signal-to-noise ratio and contrast definition on the sensitivity assessment and benchmarking of fluorescence molecular imaging systems.Journal of biomedical optics · 2025Article
- A multimodal imaging workflow to visualize metal mixtures in the human placenta and explore colocalization with biological response markers.Metallomics : integrated biometal science · 2016Article
- Article
- Enhanced loading of Fura-2/AM calcium indicator dye in adult rodent brain slices via a microfluidic oxygenator.Journal of neuroscience methods · 2013Article
- Distribution of aerosols in mouse lobes by fluorescent imaging.International journal of pharmaceutics · 2012Article
- A probabilistic cell model in background corrected image sequences for single cell analysis.Biomedical engineering online · 2010Article
- Measurement of the distribution of aerosols among mouse lobes by fluorescent imaging.Analytical biochemistry · 2010Article
- Improved cytoplasmic pH measurements in SNARF-1 stained plant cells by image processing.Plant signaling & behavior · 2010Article
- Activity correlation imaging: visualizing function and structure of neuronal populations.Biophysical journal · 2009Article
- Odor coding by modules of coherent mitral/tufted cells in the vertebrate olfactory bulb.Proceedings of the National Academy of Sciences of the United States of America · 2009Article
- Characterization of the mitochondrial inner membrane protein translocator Tim17 from Trypanosoma brucei.Molecular and biochemical parasitology · 2008Article
- Cell type-specific relationships between spiking and [Ca2+]i in neurons of the Xenopus tadpole olfactory bulb.The Journal of physiology · 2007Article
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4 authors.
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Abstract
Fluorescence imaging of bulk-stained tissue is a popular technique for monitoring the activities in a large population of cells. However, a precise quantification of such experiments is often compromised by an ambiguity of background estimation. Although, in single-cell-staining experiments, background can be measured from a neighboring nonstained region, such a region often does not exist in bulk-stained tissue. Here we describe a novel method that overcomes this problem. In contrast to previous methods, we determined the background of a given region of interest (ROI) using the information contained in the temporal dynamics of its individual pixels. Since no information outside the ROI is needed, the method can be used regardless of the staining profile in the surrounding tissue. Moreover, we extend the method to deal with background inhomogeneities within a single ROI, a problem not yet solved by any of the currently available tools. We performed computer simulations to demonstrate the accuracy of our method and give example applications in ratiometric calcium imaging of bulk-stained olfactory bulb slices. Converting the fluorescence signals into [Ca2+] gives resting values consistent with earlier single-cell staining results, and odorant-induced [Ca2+] transients can be quantitatively compared in different cells. Using these examples we show that inaccurate background subtraction introduces large errors (easily in the range of 100%) in the assessment of both resting [Ca2+] and [Ca2+] dynamics. The proposed method allows us to avoid such errors.
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