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
Само за регистроване кориснике
2019
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
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.
Кључне речи:
areas / hepatocytes / morphometry / stereology / volumesИзвор:
Microscopy Research and Technique, 2019, 82, 6, 861-871Издавач:
- Wiley, Hoboken
Финансирање / пројекти:
- Foundation for Science and Technology (FCT) of PortugalPortuguese Foundation for Science and Technology
- Fundacao para a Ciencia e a TecnologiaPortuguese Foundation for Science and Technology [UID/Multi/04423/2013]
- European Regional Development Fund (ERDF)European Union (EU) [UID/Multi/04423/2013]
- Унапређење производних капацитета шарана (Cyprinus carpio L) програмима исхране и селекције (RS-31075)
DOI: 10.1002/jemt.23228
ISSN: 1059-910X
PubMed: 30730589
WoS: 000467862000020
Scopus: 2-s2.0-85061280257
Институција/група
Poljoprivredni fakultetTY - 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 . .