ArticleHistochemistry and cell biology2003
A simpler way of comparing the labelling densities of cellular compartments illustrated using data from VPARP and LAMP-1 immunogold labelling experiments.
Article in Histochemistry and cell biology, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers, 1 of them a synthesis that pooled it.
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Who cites it
17 citing papers in PubMed, 1 synthesis or guideline pooled it.
- An official research policy statement of the American Thoracic Society/European Respiratory Society: standards for quantitative assessment of lung structure.American journal of respiratory and critical care medicine · 2010Guideline
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- Expression and distribution of three transient receptor potential vanilloid(TRPV) channel proteins in human odontoblast-like cells.Journal of molecular histology · 2017Article
- 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
- 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
- Quantitative assessment of specificity in immunoelectron microscopy.The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society · 2010Article
- A stereological perspective on placental morphology in normal and complicated pregnancies.Journal of anatomy · 2009Review
- Developments in cell biology for quantitative immunoelectron microscopy based on thin sections: a review.Histochemistry and cell biology · 2008Review
- Transmembrane domain-dependent partitioning of membrane proteins within the endoplasmic reticulum.The Journal of cell biology · 2008Article
- Important role of matrix metalloproteinase 9 in epileptogenesis.The Journal of cell biology · 2008Article
- Quantifying immunogold labelling patterns of cellular compartments when they comprise mixtures of membranes (surface-occupying) and organelles (volume-occupying).Histochemistry and cell biology · 2008Article
- Chromatin breakdown by deoxyribonuclease1 promotes acetaminophen-induced liver necrosis: an ultrastructural and histochemical study on male CD-1 mice.Histochemistry and cell biology · 2007Article
- Distribution of surfactant proteins in type II pneumocytes of newborn, 14-day old, and adult rats: an immunoelectron microscopic and stereological study.Histochemistry and cell biology · 2005Article
- News and views in Histochemistry and Cell Biology.Histochemistry and cell biology · 2004Review
- Golgi enzymes are enriched in perforated zones of golgi cisternae but are depleted in COPI vesicles.Molecular biology of the cell · 2004Article
- Applications of an efficient method for comparing immunogold labelling patterns in the same sets of compartments in different groups of cells.Histochemistry and cell biology · 2004Article
- Innovative techniques and applications in histochemistry and cell biology.Histochemistry and cell biology · 2003Review
Corrections and comments
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Authors and funding
6 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Quantitative immunoelectron microscopy of gold label in intracellular compartments often involves calculating labelling densities (LDs). These are related to antigen concentrations and usually refer gold particle counts to the sizes of compartments on sections (for example, golds per microm(2) of organelle profile area or per microm of membrane trace length). Here, we show how LD values can be estimated more simply (without estimating areas or lengths) and also how observed and expected LD values can be used to calculate a relative labelling index (RLI) for each compartment and then test statistically for preferential (non-random) labelling. For random labelling, RLI=1. Compartment size is estimated stereologically by superimposing random test points (which hit organelle profiles in proportion to their area) or test lines (which intersect membrane traces in proportion to their length). By this means, the observed LD of a compartment (LD(obs)) can be expressed simply as golds per test point (organelles) or per intersection (membranes). Furthermore, the LD obtained by dividing total golds (on all compartments) by total points or intersections (on all compartments) is the value to be expected (LD(exp)) when compartments label randomly. For each compartment, RLI=LD(obs)/LD(exp). Statistical analysis is undertaken by comparing observed distributions of golds with predicted random distributions (calculated from point or intersection counts). A compartment is preferentially labelled if two criteria are met: (1) its RLI>1 (i.e. LD(obs) is greater than LD(exp)) and (2) its partial chi-squared value makes a substantial contribution to total chi-squared value. This approach provides a simple and efficient way of comparing LDs in different compartments. Its utility is illustrated using data from VPARP and LAMP-1 labelling experiments.
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