Please use this identifier to cite or link to this item: http://hdl.handle.net/2445/101504
Title: Cryostat slice irregularities may introduce bias in tissue thickness estimation: relevance for cell counting methods
Author: Puigdellívol Sánchez, Anna
Giralt Torroella, Albert
Casanovas, A.
Alberch i Vié, Jordi
Prats Galino, Alberto
Keywords: Microscòpia mèdica
Cèl·lules
Visualització tridimensional
Medical microscopy
Cells
Three-dimensional display systems
Issue Date: 15-Jul-2015
Publisher: Cambridge University Press
Abstract: Stereological techniques using the optical disectors require estimation of final section thickness, but frozen tissue irregularities may interfere with this estimation. Cryostat slices from rodent nerve tissues (dorsal root ganglia, spinal cord, and brain), cut at 16, 40, and 50 μm, were digitized with a confocal microscope and visualized through 3D software. Geometric section thickness of tissue (Tgeom) was defined as tissue volume/area. Maximal section thicknesses (Tmax), from the top to the bottom of the section, were measured in a random sample of vertical ZX planes. Irregularities were mostly related to blood vessels traversing the tissue and neuronal somas protruding over the cut surfaces, with other neuron profiles showing a fragmented appearance. Irregularities contributed to increasing the distance between the tops and bottoms of slices sectioned in different laboratories. Significant differences were found between Tmax and Tgeom for all thickness studies and counting frames (p<0.01). The Tgeom/Tmax average rate was 68.4 85.7% in volumes around cell profiles (∼600 1,200 μm2) and 83.3 91.8% in subcellular samples (∼25 160 μm2). Confocal microscopy may help to assess tissue irregularities, which might lead to an overestimation of tissue volume if section thickness is estimated by focusing on the top and bottom of the sections.
Note: Reproducció del document publicat a: http://dx.doi.org/10.1017/S143192761501380X
It is part of: Microscopy and Microanalysis, 2015, vol. 21, num. 4
Related resource: http://dx.doi.org/10.1017/S143192761501380X
URI: http://hdl.handle.net/2445/101504
ISSN: 1431-9276
Appears in Collections:Articles publicats en revistes (Cirurgia i Especialitats Medicoquirúrgiques)

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