PEA-CLARITY: 3D molecular imaging of whole plant organs

Scientific Reports
William M PalmerChristopher P L Grof

Abstract

Here we report the adaptation of the CLARITY technique to plant tissues with addition of enzymatic degradation to improve optical clearing and facilitate antibody probe penetration. Plant-Enzyme-Assisted (PEA)-CLARITY, has allowed deep optical visualisation of stains, expressed fluorescent proteins and IgG-antibodies in Tobacco and Arabidopsis leaves. Enzyme treatment enabled penetration of antibodies into whole tissues without the need for any sectioning of the material, thus facilitating protein localisation of intact tissue in 3D whilst retaining cellular structure.

References

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Citations

Jan 7, 2016·Cell Biology International·Andrew Milgroom, Evelyn Ralston
Mar 31, 2016·Journal of Microscopy·Antonius C J Timmers
Jun 28, 2016·Microscopy Research and Technique·Katharine R MinkerJeffrey L Caplan
May 31, 2016·Molecules and Cells·Jinyoung SeoSung-Yon Kim
Jun 15, 2016·Annual Review of Cell and Developmental Biology·Kazuki TainakaHiroki R Ueda
Sep 7, 2016·Scientific Reports·Kristian H R Jensen, Rune W Berg
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Sep 7, 2018·Experimental and Therapeutic Medicine·Hao DuQiyu Li
Dec 28, 2016·Journal of Experimental Botany·Karen J I LeeEnrico S Coen
Oct 13, 2017·Journal of Biophotonics·Tingting YuDan Zhu
Feb 20, 2020·Plant Direct·Leonie H LuginbuehlJulian M Hibberd
Apr 28, 2018·Scientific Reports·Yongtai YinMaoteng Li
Jan 16, 2021·Scientific Reports·Anne FabricantDmitry Budker
Mar 16, 2021·IScience·Tingting YuDan Zhu
Jun 24, 2020·The Journal of Investigative Dermatology·Yingrou TanLai Guan Ng
Oct 2, 2021·Development·Alba Vieites-Prado, Nicolas Renier

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Methods Mentioned

BETA
confocal microscopy
imaging techniques
chemical treatments
light sheet microscopy

Software Mentioned

SeeDB
Leica Applications Suite — Fluorescence ( LAS - AF )
Leica SP8
3DISCO
Sca
CLARITY
PEA
PACT

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