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.

Základní údaje

Originální název

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

Autoři

ATTULURI, Venkata Pardha S. (356 Indie, domácí), Juan Francisco SANCHEZ LOPEZ (724 Španělsko, domácí), Lukáš MAIER (203 Česká republika, domácí), Kamil PARUCH (203 Česká republika, domácí) a Helene ROBERT BOISIVON (250 Francie, garant, domácí)

Vydání

Plant Reproduction, Springer, 2022, 2194-7953

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Kód RIV

RIV/00216224:14740/22:00127169

Organizace

Středoevropský technologický institut – Masarykova univerzita – Repozitář

DOI

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

UT WoS

000884195100001

Klíčová slova anglicky

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

Návaznosti

EF16_019/0000738, projekt VaV. EF16_025/0007381, projekt VaV. LM2018130, projekt VaV. Czech-BioImaging II, velká výzkumná infrastruktura.
Změněno: 9. 10. 2024 00:50, RNDr. Daniel Jakubík

Anotace

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.
Zobrazeno: 11. 5. 2025 06:51