Gundersen, H. J. G. et al. Some new, simple and efficient stereological methods and their use in pathological research and diagnosis. APMIS 96, 379−394

Apmis (Impact Factor: 2.04). 12/1987; 96(1‐6):379 - 394. DOI: 10.1111/j.1699-0463.1988.tb05320.x
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Stereology is a set of simple and efficient methods for quantitation of three-dimensional microscopic structures which is specifically tuned to provide reliable data from sections. Within the last few years, a number of new methods has been developed which are of special interest to pathologists. Methods for estimating the volume, surface area and length of any structure are described in this review. The principles on which stereology is based and the necessary sampling procedures are described and illustrated with examples. The necessary equipment, the measurements, and the calculations are invariably simple and easy.

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    • "Stereology requires that the region of interest is clearly identifiable and that the particles are recognizable in an unambiguous fashion (Gundersen et al. 1988; Williams and Rakic 1988; Schmitz and Hof 2005; Geuna and Herrera-Rincon 2015). Counting is performed within an Boptical disector,^ a 3D polyhedral probe placed within tissue sections and analyzed through focal planes available with high-resolution microscopy by using an Bunbiased counting scheme^ that includes all particles inside the disector and those particles that touch any of the three Binclusion surfaces^ of the probe but that ignores those that touch any of the three Bexclusion surfaces^ of the probe (Gundersen 1986; Williams and Rakic 1988; Schmitz and Hof 2005). "
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    ABSTRACT: The number of cells comprising biological structures represents fundamental information in basic anatomy, development, aging, drug tests, pathology and genetic manipulations. Obtaining unbiased estimates of cell numbers, however, was until recently possible only through stereological techniques, which require specific training, equipment, histological processing and appropriate sampling strategies applied to structures with a homogeneous distribution of cell bodies. An alternative, the isotropic fractionator (IF), became available in 2005 as a fast and inexpensive method that requires little training, no specific software and only a few materials before it can be used to quantify total numbers of neuronal and non-neuronal cells in a whole organ such as the brain or any dissectible regions thereof. This method entails transforming a highly anisotropic tissue into a homogeneous suspension of free-floating nuclei that can then be counted under the microscope or by flow cytometry and identified morphologically and immunocytochemically as neuronal or non-neuronal. We compare the advantages and disadvantages of each method and provide researchers with guidelines for choosing the best method for their particular needs. IF is as accurate as unbiased stereology and faster than stereological techniques, as it requires no elaborate histological processing or sampling paradigms, providing reliable estimates in a few days rather than many weeks. Tissue shrinkage is also not an issue, since the estimates provided are independent of tissue volume. The main disadvantage of IF, however, is that it necessarily destroys the tissue analyzed and thus provides no spatial information on the cellular composition of biological regions of interest.
    Cell and Tissue Research 03/2015; 360(1). DOI:10.1007/s00441-015-2127-6 · 3.57 Impact Factor
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    • "Unbiased estimations of the density of NeuN (n = 6 mice/ group), CR (n = 6/group), CB (n = 8/group), TH (n = 3/ group), and PV (n = 3/group) immunopositive cells were performed using an optical dissector unbiased sample design (Gundersen et al. 1988). Stereological analysis was performed with an Olympus BX51 microscope with a calibrated motorised stage controller that allows precise control of x-, y-and z-axes linked to the New CAST software. "
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    ABSTRACT: New subventricular zone (SVZ)-derived neuroblasts that migrate via the rostral migratory stream are continuously added to the olfactory bulb (OB) of the adult rodent brain. Anosmin-1 (A1) is an extracellular matrix protein that binds to FGF receptor 1 (FGFR1) to exert its biological effects. When mutated as in Kallmann syndrome patients, A1 is associated with severe OB morphogenesis defects leading to anosmia and hypogonadotropic hypogonadism. Here, we show that A1 over-expression in adult mice strongly increases proliferation in the SVZ, mainly with symmetrical divisions, and produces substantial morphological changes in the normal SVZ architecture, where we also report the presence of FGFR1 in almost all SVZ cells. Interestingly, for the first time we show FGFR1 expression in the basal body of primary cilia in neural progenitor cells. Additionally, we have found that A1 over-expression also enhances neuroblast motility, mainly through FGFR1 activity. Together, these changes lead to a selective increase in several GABAergic interneuron populations in different OB layers. These specific alterations in the OB would be sufficient to disrupt the normal processing of sensory information and consequently alter olfactory memory. In summary, this work shows that FGFR1-mediated A1 activity plays a crucial role in the continuous remodelling of the adult OB.
    Brain Structure and Function 10/2014; DOI:10.1007/s00429-014-0904-8 · 5.62 Impact Factor
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    • "The volume of the three subnuclei was calculated as the sum of their areas multiplied by the distance between the upper planes of the examined sections (Gundersen et al. 1988; Uylings et al. 1986), as determined using the measured thickness of the tissue (West et al. 1991). In the absence of evidence for differential tissue shrinkage according to group (Table 1), these post-processing tissue dimensions were used for statistical comparisons of BLA volume, without shrinkage correction. "
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    ABSTRACT: Functional imaging studies consistently report abnormal amygdala activity in major depressive disorder (MDD). Neuroanatomical correlates are less clear: imaging studies have produced mixed results on amygdala volume, and postmortem neuroanatomic studies have only examined cell densities in portions of the amygdala or its subregions in MDD. Here, we present a stereological analysis of the volume of, and the total number of, neurons, glia, and neurovascular (pericyte and endothelial) cells in the basolateral amygdala in MDD. Postmortem tissues from 13 subjects with MDD and 10 controls were examined. Sections (~15/subject) taken throughout the rostral-caudal extent of the basolateral amygdala (BLA) were stained for Nissl substance and utilized for stereological estimation of volume and cell numbers. Results indicate that depressed subjects had a larger lateral nucleus than controls and a greater number of total BLA neurovascular cells than controls. There were no differences in the number or density of neurons or glia between depressed and control subjects. These findings present a more detailed picture of BLA cellular anatomy in depression than has previously been available. Further studies are needed to determine whether the greater number of neurovascular cells in depressed subjects may be related to increased amygdala activity in depression.
    Brain Structure and Function 10/2014; DOI:10.1007/s00429-014-0900-z · 5.62 Impact Factor
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