Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Modelling the short-term response to nitrogen that coordinates events in lateral root initiation.

Quantitative plant biology·2026
Same author

Phosphate starvation induces root cell-type-specific transcriptional responses and alternative splicing.

The New phytologist·2026
Same author

Root growth promotion by <i>Penicillium melinii</i> : mechanistic insights and agricultural applications.

bioRxiv : the preprint server for biology·2026
Same author

Root and microbiome synergy in plant heat stress resilience: epigenetic regulation as a frontier for future research.

Journal of experimental botany·2025
Same author

Exogenous bacterial cellulose induces plant tissue regeneration through the regulation of cytokinin and defense networks.

Science advances·2025
Same author

Advancing our understanding of root development: Technologies and insights from diverse studies.

Plant physiology·2024

Related Experiment Video

Updated: Dec 24, 2025

Fluorescence Activated Cell Sorting of Plant Protoplasts
13:35

Fluorescence Activated Cell Sorting of Plant Protoplasts

Published on: February 18, 2010

25.4K

Fluorescence-Activated Cell Sorting Using the D-Root Device and Optimization for Scarce and/or Non-Accessible Root

Mary-Paz González-García1, Estéfano Bustillo-Avendaño1, Alvaro Sanchez-Corrionero1

  • 1Centro de Biotecnología y Genómica de Plantas (Universidad Politécnica de Madrid-Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria), Campus de Montegancedo, Pozuelo de Alarcón, 28223 Madrid, Spain.

Plants (Basel, Switzerland)
|April 17, 2020
PubMed
Summary

We developed a new Fluorescence-Activated Cell Sorting (FACS) protocol to isolate plant root cells grown in darkness. This method enables transcriptomic analysis of root cell types under natural conditions.

Keywords:
D-RootFACScell-typefounder cellslateral rootsroot protoplasts

More Related Videos

Cell Specific Analysis of Arabidopsis Leaves Using Fluorescence Activated Cell Sorting
11:25

Cell Specific Analysis of Arabidopsis Leaves Using Fluorescence Activated Cell Sorting

Published on: October 4, 2012

15.7K
Fluorescence-Activated Cell Sorting for the Isolation of Scleractinian Cell Populations
04:32

Fluorescence-Activated Cell Sorting for the Isolation of Scleractinian Cell Populations

Published on: May 31, 2020

8.5K

Related Experiment Videos

Last Updated: Dec 24, 2025

Fluorescence Activated Cell Sorting of Plant Protoplasts
13:35

Fluorescence Activated Cell Sorting of Plant Protoplasts

Published on: February 18, 2010

25.4K
Cell Specific Analysis of Arabidopsis Leaves Using Fluorescence Activated Cell Sorting
11:25

Cell Specific Analysis of Arabidopsis Leaves Using Fluorescence Activated Cell Sorting

Published on: October 4, 2012

15.7K
Fluorescence-Activated Cell Sorting for the Isolation of Scleractinian Cell Populations
04:32

Fluorescence-Activated Cell Sorting for the Isolation of Scleractinian Cell Populations

Published on: May 31, 2020

8.5K

Area of Science:

  • Plant Biology
  • Molecular Biology
  • Genomics

Background:

  • Fluorescence-activated cell sorting (FACS) is crucial for isolating specific cell populations for transcriptomic analysis.
  • Existing protoplast extraction protocols are unsuitable for root cells grown in darkness, a non-natural condition.
  • Root cells typically grow in darkness, and current methods fail to capture this under natural conditions.

Purpose of the Study:

  • To establish an optimized Fluorescence-Activated Cell Sorting (FACS) protocol for isolating root protoplasts from plants grown in darkness.
  • To enable transcriptomic analysis of specific, even scarce, root cell types under simulated natural conditions.
  • To provide a robust method for studying root cell-type specific gene expression.

Main Methods:

  • Utilized Arabidopsis green fluorescent protein (GFP)-marked lines and the D-Root device to grow roots in darkness.
  • Optimized protoplast isolation using a minimal enzyme concentration for efficient yield.
  • Employed Fluorescence-Activated Cell Sorting (FACS) with mCherry fluorescent protein for purification of specific cell types.

Main Results:

  • Successfully isolated viable root protoplasts from Arabidopsis grown in darkness using an optimized FACS protocol.
  • Demonstrated the protocol's effectiveness in isolating scarce cell types from differentiated tissues.
  • Generated cDNA and sequencing libraries from isolated RNA, confirming suitability for genome-wide transcriptomic analysis.

Conclusions:

  • The developed FACS protocol effectively isolates root protoplasts from plants grown in darkness.
  • This method allows for cell-type specific transcriptomic profiling of root cells under near-natural conditions.
  • The protocol is versatile and applicable to various Arabidopsis lines and cell types for gene expression studies.