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- Unbiased stereology using optical fractiontor probe ki67 serial#
- Unbiased stereology using optical fractiontor probe ki67 software#
Histologic sections are commonly analyzed by area morphometry or unbiased stereology, but stereology requires specialized equipment. Wakasaki, Rumie Eiwaz, Mahaba McClellan, Nicholas Matsushita, Katsuyuki Golgotiu, Kirsti Hutchens, Michael PĪ technical challenge in translational models of kidney injury is determination of the extent of cell death. In addition, volume reconstruction facilitates visualizationĪutomated systematic random sampling and Cavalieri stereology of histologic sections demonstrating acute tubular necrosis after cardiac arrest and cardiopulmonary resuscitation in the mouse. These analysis methods are less sensitive to the section-to-section variations in counts and section thickness, factors that contribute to the inaccuracy of other stereological methods. RESULTS AND CONCLUSIONS: Reconstruction makes practicable volume-oriented analysis of ultrastructure using such techniques as the unbiased bricking and fractional counter methods. A volume of brain tissue from stratum radiatum of hippocampal area CA1 is reconstructed and analyzed for synaptic density to demonstrate and compare the techniques. This unbiased technique relies on the counting of fractions of objects contained in a test volume. A new general method, the fractional counter, is also described. Stereological techniques for assessing structural distributions in reconstructed volumes are the unbiased bricking, disector, unbiased ratio, and per-length counting techniques.
Unbiased stereology using optical fractiontor probe ki67 software#
Computer software for generating transformations based on user input is described. Such transformations are implemented by a linear combination of bivariate polynomials. Whole-field alignment requires rotation, translation, skew, scaling, and second-order nonlinear deformations. DESIGN AND MEASUREMENTS: Volumes are reconstructed by defining transformations that align the entire area of adjacent sections. This paper introduces a new reconstruction system and new methods for analyzing in three dimensions the location and ultrastructure of neuronal components, such as synapses, which are distributed non-randomly throughout the brain.
Unbiased stereology using optical fractiontor probe ki67 serial#
Reconstruction from multiple serial sections provides a much needed, richer representation of the three-dimensional organization of the brain. In the past, such analyses have usually been based on single or paired sections obtained by electron microscopy.
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OBJECTIVE: Analysis of brain ultrastructure is needed to reveal how neurons communicate with one another via synapses and how disease processes alter this communication. When properly applied, fluorescent probes are ideally suited to the three-dimensional sampling requirements of the optical fractionator procedure.Extending unbiased stereology of brain ultrastructure to three-dimensional volumesįiala, J. The chapter concludes with a summary of the advantages and disadvantages of using fluorescent probes for quantitative stereology. It also describes the limitations of fluorescent probes and cautions about their analysis. It demonstrates the advantages of fluorescent probes for optical sectioning and the preservation of section thickness that results from the preparative methodology making staining with fluorescent probes particularly useful for the implementation of the optical fractionator procedure. The theoretical and practical considerations for using fluorescent probes in quantitative analysis are presented along with a worked example using the optical fractionator procedure. This chapter describes the use of fluorescent probes in cell-counting procedures. Chapter 15 Unbiased morphometrical techniques for the quantitative assessment of cells in primary dissociation cultures.Chapter 13 Virtual test systems for estimation of orientation-dependent parameters in thick, arbitrarily orientated sections exemplified by length quantification of regenerating axons in spinal cord lesions using isotropic, virtual planes.Chapter 12 Length estimation of nerve fibers in human white matter using isotropic, uniformly random sections.Chapter 10 Estimation of number and volume of immunohistochemically stained neurons in complex brain regions.Chapter 9 The nucleator and the planar and optical rotators applied in rat dorsal root ganglia.Chapter 7 The number of microvessels estimated by an unbiased stereological method applied in a brain region.Chapter 6 Number in electron microscopy: estimation of total number of synapses in the main regions of human neocortex.
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