ArticleThe journal of histochemistry and cytochemistry : official journal of the Histochemistry Society2010
Quantitative assessment of specificity in immunoelectron microscopy.
Article in The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society, 2010. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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Who cites it
12 citing papers in PubMed, 22 citations in OpenAlex.
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- Altered Basal Autophagy Affects Extracellular Vesicle Release in Cells of Lagotto Romagnolo Dogs With a VariantVeterinary pathology · 2020Article
- Quantitative basis of meiotic chromosome synapsis analyzed by electron tomography.Scientific reports · 2019Article
- Phospholipase C-η2 interacts with nuclear and cytoplasmic LIMK-1 during retinoic acid-stimulated neurite growth.Histochemistry and cell biology · 2016Article
- From gross anatomy to the nanomorphome: stereological tools provide a paradigm for advancing research in quantitative morphomics.Journal of anatomy · 2015Article
- Plasma membrane-located purine nucleotide transport proteins are key components for host exploitation by microsporidian intracellular parasites.PLoS pathogens · 2014Article
- Antibodies for immunolabeling by light and electron microscopy: not for the faint hearted.Histochemistry and cell biology · 2014Review
- Mapping the distributions and quantifying the labelling intensities of cell compartments by immunoelectron microscopy: progress towards a coherent set of methods.Journal of anatomy · 2011Review
- The cytoplasmic domain of Marburg virus GP modulates early steps of viral infection.Journal of virology · 2011Article
- Multiple-labelling immunoEM using different sizes of colloidal gold: alternative approaches to test for differential distribution and colocalization in subcellular structures.Histochemistry and cell biology · 2011Article
Corrections and comments
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Authors and funding
2 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
In immunoelectron microscopy (immuno-EM) on ultrathin sections, gold particles are used for localization of molecular components of cells. These particles are countable, and quantitative methods have been established to estimate and evaluate the density and distribution of "raw" gold particle counts from a single uncontrolled labeling experiment. However, these raw counts are composed of two distinct elements: particles that are specific (specific labeling) and particles that are not (nonspecific labeling) for the target component. So far, approaches for assessment of specific labeling and for correction of raw gold particle counts to reveal specific labeling densities and distributions have not attracted much attention. Here, we discuss experimental strategies for determining specificity in immuno-EM, and we present methods for quantitative assessment of (1) the probability that an observed gold particle is specific for the target, (2) the density of specific labeling, and (3) the distribution of specific labeling over a series of compartments. These methods should be of general utility for researchers investigating the distribution of cellular components using on-section immunogold labeling.
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