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

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.; Juan Francisco SANCHEZ LOPEZ; Lukáš MAIER; Kamil PARUCH a Helene ROBERT BOISIVON

Vydání

Plant Reproduction, Springer, 2022, 2194-7953

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10604 Reproductive biology

Stát vydavatele

Spojené státy

Utajení

není předmětem státního či obchodního tajemství

Odkazy

open access published article

Impakt faktor

Impact factor: 3.400

Označené pro přenos do RIV

Ano

Kód RIV

RIV/00216224:14740/22:00127169

Organizační jednotka

Středoevropský technologický institut

Klíčová slova anglicky

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

Štítky

14313010, 143160, CF CELLIM, CF PLANT, rivok

Příznaky

Mezinárodní význam, Recenzováno

Změněno: 8. 10. 2024 10:33, Ing. Martina Blahová

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.

Návaznosti

EF16_019/0000738, projekt VaV

Název: Centrum experimentální biologie rostlin

EF16_025/0007381, projekt VaV

Název: Preklinická progrese nových organických sloučenin s cílenou biologickou aktivitou

LM2018130, projekt VaV

Název: Národní infrastruktura chemické biologie (Akronym: CZ-OPENSCREEN)

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, CZ-OPENSCREEN: National Infrastructure for Chemical Biology

90129, velká výzkumná infrastruktura

Název: Czech-BioImaging II

Přiložené soubory

Attuluri_et_al-2022-Plant_Reproduction.pdf

Citovat

ATTULURI, Venkata Pardha S.; Juan Francisco SANCHEZ LOPEZ; Lukáš MAIER; Kamil PARUCH a Helene ROBERT BOISIVON. Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana. Plant Reproduction. Springer, 2022, roč. 35, č. 4, s. 279-293. ISSN 2194-7953. Dostupné z: https://doi.org/10.1007/s00497-022-00453-4.

@article{2232149,
   author = {Attuluri, Venkata Pardha S. and Sanchez Lopez, Juan Francisco and Maier, Lukáš and Paruch, Kamil and Robert Boisivon, Helene},
   article_number = {4},
   doi = {https://doi.org/10.1007/s00497-022-00453-4},
   keywords = {Arabidopsis seed clearing; Microscopy imaging; Fluorescence preservation; ClearSee alpha; FAST9; Iohexol},
   language = {eng},
   issn = {2194-7953},
   journal = {Plant Reproduction},
   title = {Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana},
   url = {https://doi.org/10.1007/s00497-022-00453-4},
   volume = {35},
   year = {2022}
}

TY  - JOUR
ID  - 2232149
AU  - Attuluri, Venkata Pardha S. - Sanchez Lopez, Juan Francisco - Maier, Lukáš - Paruch, Kamil - Robert Boisivon, Helene
PY  - 2022
TI  - Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana
JF  - Plant Reproduction
VL  - 35
IS  - 4
SP  - 279-293
EP  - 279-293
PB  - Springer
SN  - 21947953
KW  - Arabidopsis seed clearing
KW  - Microscopy imaging
KW  - Fluorescence preservation
KW  - ClearSee alpha
KW  - FAST9
KW  - Iohexol
UR  - https://doi.org/10.1007/s00497-022-00453-4
N2  - 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.
ER  -

ATTULURI, Venkata Pardha S.; Juan Francisco SANCHEZ LOPEZ; Lukáš MAIER; Kamil PARUCH a Helene ROBERT BOISIVON. Comparing the efficiency of six clearing methods in developing seeds of Arabidopsis thaliana. \textit{Plant Reproduction}. Springer, 2022, roč.~35, č.~4, s.~279-293. ISSN~2194-7953. Dostupné z: https://doi.org/10.1007/s00497-022-00453-4.

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