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Abstract Detail

Development and Structure

Grasso, Matthew [1], Lintilhac, Philip [1].

Microbead encapsulation of living plant protoplasts: A new tool for the handling of single plant cells.

The current understanding of plant development has been built predominantly on physiological and genetic insights. This is in part because inquiries into the role of biomechanical signaling during plant development have been limited by a lack of experimental tools. With the expansion of microfluidic technologies which can be adapted to produce novel biological research platforms there is the potential to address this problem. Here we introduce a protocol for the incorporation of individual plant protoplasts into precisely sized agarose microbeads. The method is based on the emerging field of droplet microfluidics. Living protoplasts obtained from BY-2 tobacco suspension cultures can be continuously incorporated into a stream of agarose microdroplets and then collected in cooled mineral oil as gelled microbeads. These cells can then be transferred to MS culture medium for further analysis. In this first report we show that spherical microbeads containing living protoplasts can be easily generated in quantity and that these encapsulated cells continue to grow and divide. Microbead encapsulation of protoplasts affords the opportunity to control the physical microenvironment of individual plant cells.
By capturing individual protoplasts in agarose microbeads the cells are removed from the mechanical influence of surrounding cells and captured in a reproducible isotropic microenvironment. This makes it possible to observe how single cells respond to mechanical stimulus in a context where many physical variables have been controlled or removed. With ongoing advances in hydrogel engineering and microfluidics the technique presented here has the potential to become a powerful tool for researchers in the field of plant biomechanics and ultimately may facilitate novel studies.

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1 - University of Vermont, Dept. of Plant Biology, 63 Carrigan Dr., Dept. of Plant Biology, Burlington, VT, 05405, USA


Presentation Type: Oral Paper
Session: 29, Development and Structure I
Location: 202/Savannah International Trade and Convention Center
Date: Tuesday, August 2nd, 2016
Time: 1:30 PM
Number: 29001
Abstract ID:164
Candidate for Awards:None

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