Stereology, an Unbiased Methodological Approach to Study Plant Anatomy and Cytology: Past, Present and Future

Abstract

This review presents an historical overview of stereological methods used for the quantitative evaluation of plant anatomical and cytological structures. It includes the origins of these methods up to the most recent developments such as the application of stereology based on 3D images. We focus especially on leaf, as the vast majority of studies of plant microscopic structure examine this organ. An overview of plant cell ultrastructure measurements as well as plant anatomical characteristics (e.g., plant tissue volume density, internal leaf surface area, number and mean size of mesophyll cells and chloroplast number), which were estimated by stereological methods most frequently, is presented. We emphasize the importance of proper sampling needed for unbiased measurements. Furthermore, we mention other methods used for plant morphometric studies and briefly discuss their relevance, precision, unbiasedness and efficiency in comparison with unbiased stereology. Finally, we discuss reasons for the sparse use of stereology in plant anatomy and consider the future of stereology in plant research.

Unbiased Estimation of Norway Spruce (Picea Abies L. Karst.) Chloroplast Structure: Heterogeneity Within Needle Mesophyll Under Different Irradiance and [Co2]

“…Despite the clear importance of proper sampling in measurements of plant structure, this issue is often inadequately addressed in plant studies as pointed out recently by Kubínová et al (2017). If the study aims for estimation of unbiased quantitative parameters for the whole plant organ, then SUR and stereological methods are recommended (West, 2012).…”

Shape matters: the pitfalls of analyzing mesophyll anatomy

“…Design‐based stereological methods aim to provide estimates of, for example, volume and surface area that are independent of size, shape, orientation, or distribution of the material. Such methods typically involve taking 2D samples (images) from a 3D structure according to a given sampling design and using statistical principles to estimate 3D properties such as volume and surface area (Kubínová et al , ). The greatest advantage of such techniques for analyzing the leaf mesophyll is that no assumptions on cell shapes are necessary.…”

Image Analysis & Stereology: 2017 Research Highlights

“…The special topic was continued by Kubínová et al (2017), who provided a historical overview of stereology as an unbiased methodological approach for the quantitative study of plant anatomy and cytology, with a particular focus on the evaluation of leaf structures with respect to the leaf being the most frequently studied plant organ. After providing a short history of the quantification of the plant anatomical structure, an overview of stereological methods applied in plant anatomy was given, focusing on different approaches to sampling, estimation of anatomical characteristics by plant surface area and volume density measurements, and plant cell ultrastructure measurements.…”