TY  - JOUR
AU  - Rašković, Božidar
AU  - Cruzeiro, Catarina
AU  - Poleksić, Vesna
AU  - Rocha, Eduardo
PY  - 2019
UR  - http://aspace.agrif.bg.ac.rs/handle/123456789/5117
AB  - Assessing fish liver status is common in aquaculture nutrition assays. This often implies determining hepatocytes profile areas in routine thin (5-7 mu m) histological sections. However, there are theoretical problems using planar morphometry in thin sections: inherent sampling cells biases, too small numbers of sampled cells, under/overestimation of size, measuring size as areas when cells are three-dimensional (3D) entities. The gold standard for assessing/validate cell size is stereology using thick sections (20-40 mu m). Here, we estimated the volume of hepatocytes and their nuclei by the nucleator and optical disector stereological probes (in thick sections), and, innovatively, in thin sections too (using single-section disectors). The liver of common carp eating feed containing either low or high level of lipids was targeted. Results were compared with prior profile areas from planar morphometry using thin sections, and with profile areas estimated here with the two-dimensional (2D) nucleator. Ratios between nucleus and cell/cytoplasm (N/C) areas and volumes were calculated and compared. There was high positive correlation between volumes in thin and thick sections (r = .85 to .89; p  lt  .001), empirically validating the single-section disector. Strong correlations existed between profile-derived versus 2D-nucleator areas (r = .74 to .83; p  lt  .001). There was systematic underestimation of cells and nucleus size using planar morphometry. The N/C ratios derived from the 2D-nucleator data were higher than those from planar morphometry. Despite theoretical premises for using simple planar morphometry in thin sections are flawed, our results support that such morphometry on carp/fish hepatocytes may offer some valid biological conclusions. Anyway, we advanced guidelines for implementing proper methods.
PB  - Wiley, Hoboken
T2  - Microscopy Research and Technique
T1  - Estimating volumes from common carp hepatocytes using design-based stereology and examining correlations with profile areas: Revisiting a nutritional assay and unveiling guidelines to microscopists
EP  - 871
IS  - 6
SP  - 861
VL  - 82
DO  - 10.1002/jemt.23228
ER  - 

@article{
author = "Rašković, Božidar and Cruzeiro, Catarina and Poleksić, Vesna and Rocha, Eduardo",
year = "2019",
abstract = "Assessing fish liver status is common in aquaculture nutrition assays. This often implies determining hepatocytes profile areas in routine thin (5-7 mu m) histological sections. However, there are theoretical problems using planar morphometry in thin sections: inherent sampling cells biases, too small numbers of sampled cells, under/overestimation of size, measuring size as areas when cells are three-dimensional (3D) entities. The gold standard for assessing/validate cell size is stereology using thick sections (20-40 mu m). Here, we estimated the volume of hepatocytes and their nuclei by the nucleator and optical disector stereological probes (in thick sections), and, innovatively, in thin sections too (using single-section disectors). The liver of common carp eating feed containing either low or high level of lipids was targeted. Results were compared with prior profile areas from planar morphometry using thin sections, and with profile areas estimated here with the two-dimensional (2D) nucleator. Ratios between nucleus and cell/cytoplasm (N/C) areas and volumes were calculated and compared. There was high positive correlation between volumes in thin and thick sections (r = .85 to .89; p  lt  .001), empirically validating the single-section disector. Strong correlations existed between profile-derived versus 2D-nucleator areas (r = .74 to .83; p  lt  .001). There was systematic underestimation of cells and nucleus size using planar morphometry. The N/C ratios derived from the 2D-nucleator data were higher than those from planar morphometry. Despite theoretical premises for using simple planar morphometry in thin sections are flawed, our results support that such morphometry on carp/fish hepatocytes may offer some valid biological conclusions. Anyway, we advanced guidelines for implementing proper methods.",
publisher = "Wiley, Hoboken",
journal = "Microscopy Research and Technique",
title = "Estimating volumes from common carp hepatocytes using design-based stereology and examining correlations with profile areas: Revisiting a nutritional assay and unveiling guidelines to microscopists",
pages = "871-861",
number = "6",
volume = "82",
doi = "10.1002/jemt.23228"
}

Rašković, B., Cruzeiro, C., Poleksić, V.,& Rocha, E.. (2019). Estimating volumes from common carp hepatocytes using design-based stereology and examining correlations with profile areas: Revisiting a nutritional assay and unveiling guidelines to microscopists. in Microscopy Research and Technique
Wiley, Hoboken., 82(6), 861-871.
https://doi.org/10.1002/jemt.23228

Rašković B, Cruzeiro C, Poleksić V, Rocha E. Estimating volumes from common carp hepatocytes using design-based stereology and examining correlations with profile areas: Revisiting a nutritional assay and unveiling guidelines to microscopists. in Microscopy Research and Technique. 2019;82(6):861-871.
doi:10.1002/jemt.23228 .

Rašković, Božidar, Cruzeiro, Catarina, Poleksić, Vesna, Rocha, Eduardo, "Estimating volumes from common carp hepatocytes using design-based stereology and examining correlations with profile areas: Revisiting a nutritional assay and unveiling guidelines to microscopists" in Microscopy Research and Technique, 82, no. 6 (2019):861-871,
https://doi.org/10.1002/jemt.23228 . .