The complex‐shaped ‘perforated’ synapse, a problem in quantitative stereology of the brain

Groot, Didima M.G. de; Bierman, E. P. B.

doi:10.1111/j.1365-2818.1983.tb04260.x

Cited by 32 publications

(6 citation statements)

References 18 publications

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“…With the exception of the exhaustive enumeration method 21,28 (use of which is limited by prohibitive time constraints), all can be used to estimate total podocyte number per average glomerulus.…”

Section: Concise Methodsmentioning

confidence: 99%

Estimation of Glomerular Podocyte Number

Bertram

Nicholas

White

2013

Journal of the American Society of Nephrology

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The podocyte depletion hypothesis has emerged as an important unifying concept in glomerular pathology. The estimation of podocyte number is therefore often a critical component of studies of progressive renal diseases. Despite this, there is little uniformity in the biomedical literature with regard to the methods used to estimate this important parameter. Here we review a selection of valid methods for estimating podocyte number: exhaustive enumeration method, Weibel and Gomez method, disector/Cavalieri combination, disector/fractionator combination, and thick-and-thin section method. We propose the use of the disector/fractionator method for studies in which controlled sectioning of tissue is feasible, reserving the Weibel and Gomez method for studies based on archival or routine pathology material.

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Section: Concise Methodsmentioning

confidence: 99%

Estimation of Glomerular Podocyte Number

Bertram

Nicholas

White

2013

Journal of the American Society of Nephrology

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“…These difficulties are compounded by the presence of perforated synapses, including those with a segmented PSD, in many regions of the central nervous system (Calverley & Jones, 1990). A perforated synapse can be erroneously identified in a single section as two or more nonperforated ones (De Groot & Bierman, 1983) or not identified at all if the section passes through the interval between separate PSD segments (Geinisman et al, 1987). Thus, an unambiguous identification of synapses, which is a prerequisite for obtaining unbiased estimates of their number, can only be performed with the aid of consecutive serial sections.…”

Section: Ldentifljing Synapses In Serial Rather Than In Single Ultrmentioning

confidence: 99%

Unbiased stereological estimation of the total number of synapses in a brain region

et al. 1996

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SummaryModern stereological methods have been used to make unbiased estimates of the total number of synapses in the striatum radiatum of the hippocampal CA1 region of five rabbits. The approach used involved a two stage analysis and is generally applicable to all parts of the nervous system. During the first stage of the analysis, the reference volume was estimated by point counting, at the light microscope level, according to the Cavalieri principle. During the second stage, the numerical density of synapses was estimated with disectors at the electron microscopic level. The total number of synapses was calculated as the product of the numerical density and the volume of the region. The sampling with points and disectors was carried out in all three dimensions of the entire CA1 region in a manner that ensured that all parts of the region and all synapses within it had equal probabilities of being sampled. An analysis of the precision of the estimate of total synapse number has been performed in terms of the variances of volume and synaptic numerical density at different levels of sampling, i.e. at the level of points, sections, individual animals and group of animals. Detailed descriptions of the procedures used to estimate the total number of synapses, evaluate the precision of the estimates, and optimize the sampling scheme are provided.

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“…In previous studies (De Groot & Bierman, 1983, 1986, 1988De Groot, 1984 the 'small-fold' method (Small, 1968) has been used to estimate t ' . In these studies the numerical density of complex-shaped (perforated) synapses was estimated and their shape reconstructed, using serial sections (CNZ Orive, 1980De Groot & Bierman, 1983,1986,1988De Groot, 1984;Sterio, 1984). The small-fold method, however, cannot be directly adapted since the tracking of the investigated particles through the series of sections would be hampered by the presence of folds in the sections, a prerequisite for the small-fold method.…”

Section: Introductionmentioning

confidence: 99%

Comparison of methods for the estimation of the thickness of ultrathin tissue sections

Groot

1988

Journal of Microscopy

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For a number of quantitative electron microscopical techniques it is relevant to obtain an estimate of the thickness t' of the section which, in general, will differ from the actual distance t between the two cuts that generate the section. To estimate t' of ultrathin sections, several techniques have been adopted in the past, both with and without the aid of the electron microscope and additional equipment, which are summarized in an appendix. In the present study five methods have been evaluated experimentally using sections of ten different interference colours: (a) the 'small-fold' technique, (b) the 'electron scattering' method, (c) interference microscopy with (A) the Vicker's M86@ scanning microinterferometer and (B) the Jenoptik Amplival Interphako@ interference microscope and (d) the 're-embedding' method. Reliable, reproducible and comparable results were obtained with the small-fold technique, with the Vickers M86@ scanning microinterferometer and with the electron scattering method. For the last method, standard test lines for the different settings of the electron microscope were developed. The results obtained with the Jenoptik Amplival InterphakoB interference microscope are reproducible, but show a constant difference, i.e. a factor of 1-36, in thickness compared with the other three techniques. The possible cause of this 'systematic error' is discussed. The re-embedding method proved to be more laborious and slightly less reliable than the other techniques.The variation in t' between sections of a particular interference colour (inter-section variation) was found to be larger than the variation in t' within a section (intra-section variation).

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The complex‐shaped ‘perforated’ synapse, a problem in quantitative stereology of the brain

Cited by 32 publications

References 18 publications

Estimation of Glomerular Podocyte Number

Estimation of Glomerular Podocyte Number

Unbiased stereological estimation of the total number of synapses in a brain region

Comparison of methods for the estimation of the thickness of ultrathin tissue sections

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