J 2022

Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana

ATTULURI, Venkata Pardha S., Juan Francisco SANCHEZ LOPEZ, Lukáš MAIER, Kamil PARUCH, Helene ROBERT BOISIVON et. al.

Basic information

Original name

Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana

Authors

ATTULURI, Venkata Pardha S. (356 India, belonging to the institution), Juan Francisco SANCHEZ LOPEZ (724 Spain, belonging to the institution), Lukáš MAIER (203 Czech Republic, belonging to the institution), Kamil PARUCH (203 Czech Republic, belonging to the institution) and Helene ROBERT BOISIVON (250 France, guarantor, belonging to the institution)

Edition

Plant Reproduction, Springer, 2022, 2194-7953

Other information

Language

English

Type of outcome

Article in a journal

Country of publisher

United States of America

Confidentiality degree

is not subject to a state or trade secret

References:

URL

RIV identification code

RIV/00216224:14740/22:00127169

Organization

Středoevropský technologický institut – Repository – Repository

DOI

http://dx.doi.org/10.1007/s00497-022-00453-4

UT WoS

000884195100001

Keywords in English

Arabidopsis seed clearing; Microscopy imaging; Fluorescence preservation; ClearSee alpha; FAST9; Iohexol

Links

EF16_019/0000738, research and development project. EF16_025/0007381, research and development project. LM2018130, research and development project. Czech-BioImaging II, large research infrastructures.
Changed: 9/10/2024 00:50, RNDr. Daniel Jakubík

Abstract

V originále

Tissue clearing methods eliminate the need for sectioning, thereby helping better understand the 3D organization of tissues and organs. In the past fifteen years, clearing methods have been developed to preserve endogenous fluorescent protein tags. Some of these methods (ClearSee, TDE, PEA-Clarity, etc.) were adapted to clear various plant species, with the focus on roots, leaves, shoot apical meristems, and floral parts. However, these methods have not been used in developing seeds beyond the early globular stage. Tissue clearing is problematic in post-globular seeds due to various apoplastic barriers and secondary metabolites. In this study, we compared six methods for their efficiency in clearing Arabidopsis thaliana seeds at post-globular embryonic stages. Three methods (TDE, ClearSee, and ClearSee alpha) have already been reported in plants, whereas the others (fsDISCO, FAST9, and CHAPS clear) are used in this context for the first time. These methods were assessed for seed morphological changes, clearing capacity, removal of tannins, and spectral properties. We tested each method in seeds from globular to mature stages. The pros and cons of each method are listed herein. ClearSee alpha appears to be the method of choice as it preserves seed morphology and prevents tannin oxidation. However, FAST9 with 60% iohexol as a mounting medium is faster, clears better, and appears suitable for embryonic shape imaging. Our results may guide plant researchers to choose a suitable method for imaging fluorescent protein-labeled embryos in intact Arabidopsis seeds.
Displayed: 11/5/2025 06:14