The Most Comprehensive Source of Stereology Information on the Web

Stereology Information for the Biological Sciences

This site introduces both basic and advanced concepts of Stereology. The emphasis is on the use of stereology in biological research, though anyone interested in learning concepts of stereology will find something of interest. The application of stereological methods to biological studies permits researchers to effectively and efficiently gather unbiased, accurate data. This site is dedicated to helping researchers understand the principles of design-based stereology and its advantages over less sophisticated approaches in quantitative histology.

What is Stereology?

Design-based stereology is a set of methods to ensure rigorous quantitative analysis of the size, shape, and number of objects. When properly used, stereology plays an important role in validating and rejecting experimental hypotheses in biological research. It produces results that are unbiased, efficient, and more reliable than other ad hoc quantitative analyses. Unbiased stereology provides an important contribution to the advancement in biological research by improving the consistency and dependability of quantitative analytical results produced in the laboratory and reported in scientific publications.

An Introduction to Stereology Probes

The most modern unbiased stereology probes are used to quantify aspects of biological tissue in a reproducible and efficient manner. These stereological probes are appropriate for many fields of basic and applied biological and medical research. These probes should always be used in conjunction with systematic random sampling.

OPTICAL FRACTIONATOR to estimate NUMBER of cells

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Estimate the size of cell populations with the optical fractionator in thick tissue sections. In this probe, sub-volumes are sampled and then are extrapolated to arrive at an estimate of the entire cell population. A virtual space called an optical disector is used in thick sections that can be oriented anyway you like. Disector counting rules are followed to avoid overestimating, and an oil objective lens is employed for imaging, since fine z-resolution is needed to find the leading edge of the cell and to have enough focal planes to determine if it is in the disector. Note: avoid counting pieces of cells when you really want to count whole cells.    OPTICAL FRACTIONATOR

 

NUCLEATOR to estimate VOLUME of cells

Estimate individual cell volumnucleator-2es with the nucleator. A point in the cell is identified, then one to four rays are marked and their mean length is used in the formula for the volume of a sphere, generating an estimate of the cell volume. The volume estimate is number-weighted; the sampling is done with a disector in thick sections so that it is not more likely to sample larger cells than smaller cells. It is important to use a method to select cells without bias by picking them in a manner that does not favor any position of the cell in space; you won’t overestimate by sampling too many larger cross-sections or underestimate by sampling too many smaller cross-sections.    NUCLEATOR

 

CAVALIERI POINT-COUNTING to estimate VOLUME of regions

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This probe is versatile in that it can be used on thin optical or physical sections that are oriented according to the preference of the researcher. A fraction of the tissue is marked with points over the region(s) of interest, and an extrapolation is carried out to arrive at the estimate. Unbiased estimates of regional volume are easy to obtain efficiently using Cavalieri point-counting.     CAVALIERI/POINT-COUNTING

 

SPACEBALLS to estimate LENGTH of fibers and vessels

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Estimate length of biological filaments such as axons or blood vessels without facing the herculean task of tracing them all. Instead the intersections of the filaments with a virtual sphere are marked and converted to a length estimate. Thick sections may be oriented as the researcher prefers. This probe is implemented along with the fractionator method; a volume-fraction is sampled and extrapolated using the reciprocal of the volume fraction to arrive at the estimate of length.    SPACEBALLS

 

ISOTROPIC FAKIR to estimate SURFACE of membranes

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To estimate surface area, a triplet of line segments that has the property of being isotropic in space is used to probe in thick sections. Intersections of the triplet with the surface are counted and a formula is used to arrive at an estimate of the surface area; the more intersections recorded the greater the surface. Like the other regional probes on this page, thick, preferentially oriented sections are used and the fractionator method works to make an extrapolation based on the fraction of volume that was sampled.     ISOTROPIC FAKIR

more probes including solutions for thin tissue sections

 

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Recent Noteworthy Stereology Publications

Published papers using unbiased stereology; last updated on 12.13.2018.

starindicates that the researchers did a good job reporting stereological parameters. To see what parameters should be reported click here.

Predator stress-induced depression is associated with inhibition of hippocampal neurogenesis in adult male mice

starComparison of the volume fraction values of grey matter on the cervical enlargement of the spinal cord in chicken and quail

Development and biochemical characterization of a mouse model of Parkinson’s disease bearing defective glucocerebrosidase activity

Chronic Unpredictable Stress Reduces Immunostaining for Connexins 43 and 30 and Myelin Basic Protein in the Rat Prelimbic and Orbitofrontal Cortices

Fornix deep brain stimulation induces reduction of hippocampal synaptophysin levels

starGenetic inactivation of hypoxia inducible factor 1-alpha (HIF-1α) in adult hippocampal progenitors impairs neurogenesis and pattern discrimination learning

Anle138b Modulates α -Synuclein Oligomerization and Prevents Motor Decline and Neurodegeneration in a Mouse Model of Multiple System Atrophy

Non-invasive dynamic assessment of conjunctival melanomas by photoacoustic imaging

Decreased Neuron Density and Increased Glia Density in the Ventromedial Prefrontal Cortex (Brodmann Area 25) in Williams Syndrome

Vasoactive Intestinal Peptide Decreases β-Amyloid Accumulation and Prevents Brain Atrophy in the 5xFAD Mouse Model of Alzheimer’s Disease

Age‐dependent release of high‐mobility group box protein‐1 (HMGB1) and cellular neuroinflammation after traumatic brain injury in mice

Severe-combined immunodeficient rats can be used to generate a model of perinatal hypoxic-ischemic brain injury to facilitate studies of engrafted human neural stem cells.

Rush desensitization with a single antigen induces subclinical activation of mast cells and protects against bystander challenge in dually sensitized mice

Opposite development of short- and long-range anterior cingulate pathways in autism

Nigrostriatal pathology with reduced astrocytes in LRRK2 S910/S935 phosphorylation deficient knockin mice

Visualization of the Breakdown of the Axonal Transport Machinery: a Comparative Ultrastructural and Immunohistochemical Approach

Neuroprotective effects on the morphology of somatic motoneurons following the death of neighboring motoneurons: A role for microglia?

Spinal motor neurons and motor function in older adults

Reducing age-dependent monocyte-derived macrophage activation contributes to the therapeutic efficacy of NADPH oxidase inhibition in spinal cord injury

Non-invasive dynamic assessment of conjunctival melanomas using photoacoustic imaging

Maternal Preconception Stress Alters Prefrontal Cortex Development in Long–Evans Rat Pups without Changing Maternal Care

Sex- and region-specific differences in microglia phenotype and characterization of the peripheral immune response following early-life infection in neonatal male and female rats

Continue reading “Recent Noteworthy Stereology publications”

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What’s New in Stereology.info

Watch the latest webinar on Thin tissue section images with Stereo Investigator – Whole Slide Edition
Drs. Dan Peruzzi and Nathan O’Connor introduce our brand-new software, Stereo Investigator – Whole Slide Edition, designed specifically to perform unbiased stereology on large, whole slide images, and demonstrate probes. Added on 09.27.2019

Read recently published papers using unbiased stereology updated on 12.10.2018

New page: Spinal Cord Injury added on 02.09.2018

New page: Blood Vessel Pathology in a Model of Alzheimer Disease Revealed Using Space Balls added on 02.09.2018

Stereology discussion forum added

Watch the latest webinar on Using the Optical Fractionator Probe to Estimate Number of Cells with Dr. Dan Peruzzi
Learn how to count cells in an unbiased manner with the Optical Fractionator probe in Stereo Investigator. Added on: 04.24.2017

 

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