ArticlePloS one2016
A Versatile Optical Clearing Protocol for Deep Tissue Imaging of Fluorescent Proteins in Arabidopsis thaliana.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed, 49 citations in OpenAlex.
- Imaging and genetic toolbox to study Arabidopsis embryogenesis.The New phytologist · 2026Review
- Application of HEMA-AAm copolymer to achieve faster optical tissue transparency for 2D/3D fluorescence imaging.Scientific reports · 2025Article
- CsWRKY29, a key transcription factor in tea plant for freezing tolerance, ABA sensitivity, and sugar metabolism.Scientific reports · 2024Article
- Auxin response factor 10 insensitive to miR160regulation induces apospory-like phenotypes iniScience · 2024Article
- Clearing techniques for deeper imaging of plants and plant-microbe interactions.Applied microscopy · 2024Review
- Oat species and interspecific amphiploids show predominance of diploid nuclei in the syncytial endosperm.Journal of applied genetics · 2024Article
- Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana.Plant reproduction · 2022Article
- Improved clearing method contributes to deep imaging of plant organs.Communications biology · 2022Article
- ClearSeeAlpha: Advanced Optical Clearing for Whole-Plant Imaging.Plant & cell physiology · 2021Article
- A biosensor for the direct visualization of auxin.Nature · 2021Article
- Three-Dimensional Imaging in Stem Cell-Based Researches.Frontiers in veterinary science · 2021Review
- Direct fluorescence imaging of lignocellulosic and suberized cell walls in roots and stems.AoB PLANTS · 2020Article
- Seed coat development in explosively dispersed seeds of Cardamine hirsuta.Annals of botany · 2020Article
- Vibratome Sectioning and Clearing for Easing Studies of Cassava Embryo Formation.Frontiers in plant science · 2020Article
- Genetic Screens to Target Embryo and Endosperm Pathways in Arabidopsis and Maize.Methods in molecular biology (Clifton, N.J.) · 2020Article
- Anchorene is a carotenoid-derived regulatory metabolite required for anchor root formation inScience advances · 2019Article
- Constitutive signaling activity of a receptor-associated protein links fertilization with embryonic patterning inProceedings of the National Academy of Sciences of the United States of America · 2019Article
- Gibberellins negatively regulate the development of Medicago truncatula root system.Scientific reports · 2019Article
- Protocol for rapid clearing and staining of fixed Arabidopsis ovules for improved imaging by confocal laser scanning microscopy.Plant methods · 2019Article
- Multicolour three dimensional structured illumination microscopy of immunolabeled plant microtubules and associated proteins.Plant methods · 2019Article
Corrections and comments
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Authors and funding
4 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Confocal microscopy is widely used to visualize gene expression patterns and developmental processes in plants. However, the imaging of plant tissue can be challenging due to its opacity, which often makes previous immersion in a clearing agent necessary. Many commonly-used chemicals suffer either from their incompatibility with fluorescent proteins or their complex and lengthy application. 2,2'-thiodiethanol (TDE) has recently been described as a clearing agent with an emphasis on high resolution microscopy due to its potential to adjust the refractive index. Here, we evaluate the use of TDE-based clearing for confocal as well as two-photon microscopy in various Arabidopsis thaliana tissue types. We demonstrate that tissue fixation is a mandatory prerequisite for the use of TDE, in order to preserve tissue integrity and fluorescent protein activity. TDE concentrations between 50-70% are a good compromise for imaging of technically challenging tissue to achieve good clearing without affecting fluorescent protein activity. TDE-based clearing is simple and rapid to use and allows for a flexible experimental setup while facilitating high quality imaging.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.