Cargando…

Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach

Chloroplast number per cell is a frequently examined quantitative anatomical parameter, often estimated by counting chloroplast profiles in two-dimensional (2D) sections of mesophyll cells. However, a mesophyll cell is a three-dimensional (3D) structure and this has to be taken into account when qua...

Descripción completa

Detalles Bibliográficos
Autores principales: Kubínová, Zuzana, Janáček, Jiří, Lhotáková, Zuzana, Kubínová, Lucie, Albrechtová, Jana
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3904715/
https://www.ncbi.nlm.nih.gov/pubmed/24336344
http://dx.doi.org/10.1093/jxb/ert407
_version_ 1782301229786857472
author Kubínová, Zuzana
Janáček, Jiří
Lhotáková, Zuzana
Kubínová, Lucie
Albrechtová, Jana
author_facet Kubínová, Zuzana
Janáček, Jiří
Lhotáková, Zuzana
Kubínová, Lucie
Albrechtová, Jana
author_sort Kubínová, Zuzana
collection PubMed
description Chloroplast number per cell is a frequently examined quantitative anatomical parameter, often estimated by counting chloroplast profiles in two-dimensional (2D) sections of mesophyll cells. However, a mesophyll cell is a three-dimensional (3D) structure and this has to be taken into account when quantifying its internal structure. We compared 2D and 3D approaches to chloroplast counting from different points of view: (i) in practical measurements of mesophyll cells of Norway spruce needles, (ii) in a 3D model of a mesophyll cell with chloroplasts, and (iii) using a theoretical analysis. We applied, for the first time, the stereological method of an optical disector based on counting chloroplasts in stacks of spruce needle optical cross-sections acquired by confocal laser-scanning microscopy. This estimate was compared with counting chloroplast profiles in 2D sections from the same stacks of sections. Comparing practical measurements of mesophyll cells, calculations performed in a 3D model of a cell with chloroplasts as well as a theoretical analysis showed that the 2D approach yielded biased results, while the underestimation could be up to 10-fold. We proved that the frequently used method for counting chloroplasts in a mesophyll cell by counting their profiles in 2D sections did not give correct results. We concluded that the present disector method can be efficiently used for unbiased estimation of chloroplast number per mesophyll cell. This should be the method of choice, especially in coniferous needles and leaves with mesophyll cells with lignified cell walls where maceration methods are difficult or impossible to use.
format Online
Article
Text
id pubmed-3904715
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-39047152014-01-28 Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach Kubínová, Zuzana Janáček, Jiří Lhotáková, Zuzana Kubínová, Lucie Albrechtová, Jana J Exp Bot Research Paper Chloroplast number per cell is a frequently examined quantitative anatomical parameter, often estimated by counting chloroplast profiles in two-dimensional (2D) sections of mesophyll cells. However, a mesophyll cell is a three-dimensional (3D) structure and this has to be taken into account when quantifying its internal structure. We compared 2D and 3D approaches to chloroplast counting from different points of view: (i) in practical measurements of mesophyll cells of Norway spruce needles, (ii) in a 3D model of a mesophyll cell with chloroplasts, and (iii) using a theoretical analysis. We applied, for the first time, the stereological method of an optical disector based on counting chloroplasts in stacks of spruce needle optical cross-sections acquired by confocal laser-scanning microscopy. This estimate was compared with counting chloroplast profiles in 2D sections from the same stacks of sections. Comparing practical measurements of mesophyll cells, calculations performed in a 3D model of a cell with chloroplasts as well as a theoretical analysis showed that the 2D approach yielded biased results, while the underestimation could be up to 10-fold. We proved that the frequently used method for counting chloroplasts in a mesophyll cell by counting their profiles in 2D sections did not give correct results. We concluded that the present disector method can be efficiently used for unbiased estimation of chloroplast number per mesophyll cell. This should be the method of choice, especially in coniferous needles and leaves with mesophyll cells with lignified cell walls where maceration methods are difficult or impossible to use. Oxford University Press 2014-02 2013-12-11 /pmc/articles/PMC3904715/ /pubmed/24336344 http://dx.doi.org/10.1093/jxb/ert407 Text en © The Author 2013. Published by Oxford University Press on behalf of the Society for Experimental Biology. http://creativecommons.org/licenses/by/3.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Paper
Kubínová, Zuzana
Janáček, Jiří
Lhotáková, Zuzana
Kubínová, Lucie
Albrechtová, Jana
Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
title Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
title_full Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
title_fullStr Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
title_full_unstemmed Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
title_short Unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
title_sort unbiased estimation of chloroplast number in mesophyll cells: advantage of a genuine three-dimensional approach
topic Research Paper
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3904715/
https://www.ncbi.nlm.nih.gov/pubmed/24336344
http://dx.doi.org/10.1093/jxb/ert407
work_keys_str_mv AT kubinovazuzana unbiasedestimationofchloroplastnumberinmesophyllcellsadvantageofagenuinethreedimensionalapproach
AT janacekjiri unbiasedestimationofchloroplastnumberinmesophyllcellsadvantageofagenuinethreedimensionalapproach
AT lhotakovazuzana unbiasedestimationofchloroplastnumberinmesophyllcellsadvantageofagenuinethreedimensionalapproach
AT kubinovalucie unbiasedestimationofchloroplastnumberinmesophyllcellsadvantageofagenuinethreedimensionalapproach
AT albrechtovajana unbiasedestimationofchloroplastnumberinmesophyllcellsadvantageofagenuinethreedimensionalapproach