Design of a comprehensive microfluidic and microscopic toolbox for the ultra-wide spatio-temporal study of plant protoplasts development and physiology

Plant Methods
Kaori SakaiJacques Fattaccioli

Abstract

Plant protoplasts are basic plant cells units in which the pecto-cellulosic cell wall has been removed, but the plasma membrane is intact. One of the main features of plant cells is their strong plasticity, and their propensity to regenerate an organism from a single cell. Methods and differentiation protocols used in plant physiology and biology usually involve macroscopic vessels and containers that make difficult, for example, to follow the fate of the same protoplast all along its full development cycle, but also to perform continuous studies of the influence of various gradients in this context. These limits have hampered the precise study of regeneration processes. Herein, we present the design of a comprehensive, physiologically relevant, easy-to-use and low-cost microfluidic and microscopic setup for the monitoring of Physcomitrella patens (P. patens) growth and development on a long-term basis. The experimental solution we developed is made of two parts (i) a microfluidic chip composed of a single layer of about a hundred flow-through microfluidic traps for the immobilization of protoplasts, and (ii) a low-cost, light-controlled, custom-made microscope allowing the continuous recording of the moss development in physio...Continue Reading

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Citations

Mar 3, 2020·Annual Review of Plant Biology·Natalie M ClarkRosangela Sozzani
Apr 2, 2021·Frontiers in Bioengineering and Biotechnology·Rosa SagitaKristina Haslinger
May 25, 2021·Plant & Cell Physiology·Naoki YanagisawaTetsuya Higashiyama
Sep 30, 2021·The Plant Cell·Leia ColinYvon Jaillais

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Methods Mentioned

BETA
chip
chips
confocal microscopy
biopsy
2

Software Mentioned

Image J
Arduino
Kymograph Builder
Matlab
Fiji
Micro
R
ImageJ
- Manager
Python

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