Imaging plant cells and organs with light-sheet and super-resolution microscopy

Abstract

The documentation of plant growth and development requires integrative and scalable approaches to investigate and spatiotemporally resolve various dynamic processes at different levels of plant body organization. The present update deals with vigorous developments in mesoscopy, microscopy and nanoscopy methods that have been translated to imaging of plant subcellular compartments, cells, tissues and organs over the past 3 years with the aim to report recent applications and reasonable expectations from current light-sheet fluorescence microscopy (LSFM) and super-resolution microscopy (SRM) modalities. Moreover, the shortcomings and limitations of existing LSFM and SRM are discussed, particularly for their ability to accommodate plant samples and regarding their documentation potential considering spherical aberrations or temporal restrictions prohibiting the dynamic recording of fast cellular processes at the three dimensions. For a more comprehensive description, advances in living or fixed sample preparation methods are also included, supported by an overview of developments in labeling strategies successfully applied in plants. These strategies are practically documented by current applications employing model plant Arabidopsis thaliana (L.) Heynh., but also robust crop species such as Medicago sativa L. and Hordeum vulgare L. Over the past few years, the trend towards designing of integrative microscopic modalities has become apparent and it is expected that in the near future LSFM and SRM will be bridged to achieve broader multiscale plant imaging with a single platform.

Genetic manipulation of stress-induced mitogen-activated protein kinase modulates early stages of the nodulation process inMedicago sativa

“…We have recently established light-sheet fluorescence microscopy for the spatiotemporal imaging of plant development at subcellular, cellular, tissue and organ levels under controlled environmental conditions (Ovečka et al ., 2018). It was utilized in alfalfa for the characterization of root development (Vyplelová et al ., 2018) and the involvement of actin cytoskeleton in the interaction with S. meliloti (Ovečka et al ., 2022). Live-cell imaging using LSFM clearly showed relocation of SIMK from root hair tips to the S. meliloti docking site and further close association with sites of rhizobia internalization.…”

The Cytoskeleton in Plant Immunity: Dynamics, Regulation, and Function

“…Recently, Vernet et al successfully observed the nematode–root interactions in tomato using light sheet fluorescence microscopy (LSFM) and optical projection tomography (OPT) [ 117 ]. Moreover, simultaneous imaging of S. meliloti (magenta) and the host cytoskeleton in cytoskeletal marker gene transgenic alfalfa plants was realized by using light-sheet and super-resolution microscopy, providing an approach to track the dynamic changes in the host cytoskeleton and pathogenic microbes over a long time at the super-resolution level [ 118 ].…”

Plant cytokinesis and the construction of new cell wall

“…Advanced imaging technologies, such as Lattice Light Sheet (LLS) microscopy, enables long term 4D imaging without photobleaching [109] which has been used to analyse mitotic cells in mammalian cells [110] and is a promising approach for plants. Further, Light Sheet [111] and vertical imaging [112] are gaining more traction in plants and can provide cell division patterns that take the effect of gravity into account. Structured illumination microscopy has been instrumental to better understand CESA dynamics at the PM [113].…”