Stereological Principles and Sampling Procedures for Toxicologic Pathologists

Gundersen, Hans Jørgen G.; Mirabile, Rosanna C.; Brown, Danielle L.; Boyce, Rogely

doi:10.1016/b978-0-12-415759-0.00008-x

Cited by 28 publications

(88 citation statements)

References 28 publications

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“…For large organs, subsampling will need to be performed, and if paraffin processing is planned, special sampling designs have to be used to deal with shrinkage of subsamples during paraffin processing. The reader is referred to Gundersen et al (2013), where numerous sampling designs are presented that account for paraffin processing for large organs where subsampling is required.…”

Section: Automatic Proportionator Estimator: Practical Considerationsmentioning

confidence: 99%

“…It is recommended sections are first placed on a room temperature water bath to collect section pairs, then transferred on uncharged slides to a warm water bath to briefly allow sections to relax, then picked up on charged slides. Section preparation is discussed in more detail in Gundersen et al (2013). …”

Section: Automatic Proportionator Estimator: Practical Considerationsmentioning

confidence: 99%

“…The counting of polynucleated cells requires very special counting rules that are discussed in more detail in Gundersen et al (2013). …”

Section: Automatic Proportionator Estimator: Practical Considerationsmentioning

confidence: 99%

“…The question is how small? The simple answer is provided by the general inequality applicable to each group of the study (and very useful for all stereological estimators) is estimation of the Precision Range of an Optimally Balanced Estimator (PROBE) (Gundersen et al, 2013):…”

Section: Example Of Application Of the Automated Proportionator: Pilomentioning

confidence: 99%

“…To illustrate the power of the automatic proportionator estimator as a stereological tool, estimation of total number of a very rare and sparsely distributed population of osteoprogenitor cells in mouse vertebrae where total number ordinarily is below 50 cells/vertebrae was performed. For in-depth presentation of the theory of the proportionator estimator, the reader is referred to additional references (Gardi et al, 2008; Gundersen et al, 2013). …”

Section: Introductionmentioning

confidence: 99%

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The Automatic Proportionator Estimator Is Highly Efficient for Estimation of Total Number of Sparse Cell Populations

Boyce

Gundersen

2018

Front. Neuroanat.

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Estimation of total number of a population of cells that are sparsely distributed in an organ or anatomically-defined region of interest represents a challenge for conventional stereological methods. In these situations, classic fractionator approaches that rely on systematic uniform random sampling are highly inefficient and, in many cases, impractical due to the intense sampling of the organ and tissue sections that is required to obtain sufficient counts for an acceptable level of precision. The proportionator, an estimator based on non-uniform sampling theory, marries automated image analysis with stereological principles and is the only estimator that provides a highly efficient and precise method to address these challenging quantification problems. In this paper, the practical considerations of the proportionator estimator and its implementation with Proportionator™ software and digital slide imaging are reviewed. The power of the proportionator as a stereological tool is illustrated in its application to the estimation of the total number of a very rare (~50/vertebrae) and sparsely distributed population of osteoprogenitor cells in mouse vertebral body. The proportionator offers a solution to neuroscientists interested in quantifying total cell number of sparse cell populations in the central and peripheral nervous system where systematic uniform random sampling-based stereological estimators are impractical.

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Section: Automatic Proportionator Estimator: Practical Considerationsmentioning

confidence: 99%

Section: Automatic Proportionator Estimator: Practical Considerationsmentioning

confidence: 99%

“…The counting of polynucleated cells requires very special counting rules that are discussed in more detail in Gundersen et al (2013). …”

Section: Automatic Proportionator Estimator: Practical Considerationsmentioning

confidence: 99%

Section: Example Of Application Of the Automated Proportionator: Pilomentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

The Automatic Proportionator Estimator Is Highly Efficient for Estimation of Total Number of Sparse Cell Populations

Boyce

Gundersen

2018

Front. Neuroanat.

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Comparisons of Stereological and Other Approaches for Quantifying Macrophage Aggregates in Piscine Spleens

2019

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Macrophage aggregates (MAs) are focal accumulations of pigmented macrophages in the spleen and other tissues of fish. A central role of MAs is the clearance and destruction of degenerating cells and recycling of some cellular components. Macrophage aggregates also respond to chemical contaminants and infectious agents and may play a role in the adaptive immune response. Tissue damage or physiological stress can result in increased MA accumulation. As a result, MAs may be sensitive biomarkers of environmental stress in fish. Abundance of MAs in tissues has been reported in a variety of ways—most commonly as density, mean size, and relative area—but the utility of these estimates has not been compared. In this study, four different types of splenic MA abundance estimates (abundance score, density, relative area, and total volume) were compared in two fish populations (Striped Bass Morone saxatilis and White Perch M. americana) with a wide range in ages. Stereological estimates of total volume indicated an increase in MA abundance with spleen volume, which generally corresponded to fish age, and with splenic infections (mycobacteria or trematode parasites). Abundance scores were generally limited in the ability to detect changes in MA abundance by these factors, whereas density estimates were greatly influenced by changes in spleen volume. In some instances, densities declined while the total volume of MAs and spleen volume increased. Experimentally induced acute stress resulted in a decrease in spleen volume and an increase in MA density, although the total volume of MAs remained unchanged. Relative area estimates accounted for the size and number of MAs but not for changes in organ volume. Total volume is an absolute measure of MA abundance irrespective of changes in organ volume or patterns of accumulation and may provide an improved means of quantifying MAs in the spleens of fish.

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Three‐dimensional morphometry of kidney in New Zealand rabbit using unbiased design‐based stereology

Sadeghinezhad,

Lazzarini,

Bojarzadeh

et al. 2024

Microscopy Res & Technique

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The rabbit is widely used as a laboratory animal in experimental models of kidney diseases. This species is also important from a veterinary perspective as a companion animal. Stereology has been accepted as an accurate approach to kidney morphometry. The objective of the present project was to provide normal quantitative stereological parameters for adult rabbit kidneys. The left kidneys of five adult male New Zealand rabbits were used. Isotropic sections were obtained using the orientation method. Total kidney volume was calculated by the Cavalieri principle. The volume fraction of the renal structures was estimated using the point counting system. The lengths of the proximal convoluted tubule (PCT) and distal convoluted tubule (DCT) were calculated using counting frames. The total glomerular number was accounted for using the physical/fractionator technique. The mean glomerular volume was obtained by dividing the total volume of glomeruli by their total number. The total volume of rabbit kidneys calculated was 10.39 ± 1.98 cm3. The fractional volume of the kidney cortex and medulla accounted for 57.79 ± 0.65% and 42.2 ± 0.65%, respectively. The total glomerular volume was 2.18 ± 0.32% of the whole kidney. The total number of glomeruli in the rabbit kidney was estimated as 204.68 ± 12 × 103. The mean glomerular volume measured 1.07 ± 0.12 × 106 μm3. The total length of PCT and DCT was 2.96 ± 0.29 km and 1.38 ± 0.24 km, respectively. These findings can be used as a reference in experimental nephrology research and may help to expand the knowledge of nephrology in mammals by comparing with available data on humans and other species.Research Highlights Three‐dimensional morphometry of adult rabbit kidney structures was analyzed using quantitative stereology. Total volume of kidney, fractional volume of cortex and medulla, length of renal tubules and number of nephrons were estimated. These three‐dimensional morphometrical data can be used as a reference in experimental nephrology research and may help to expand the knowledge of nephrology in mammals.

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Stereological Principles and Sampling Procedures for Toxicologic Pathologists

Cited by 28 publications

References 28 publications

The Automatic Proportionator Estimator Is Highly Efficient for Estimation of Total Number of Sparse Cell Populations

The Automatic Proportionator Estimator Is Highly Efficient for Estimation of Total Number of Sparse Cell Populations

Comparisons of Stereological and Other Approaches for Quantifying Macrophage Aggregates in Piscine Spleens

Three‐dimensional morphometry of kidney in New Zealand rabbit using unbiased design‐based stereology

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