Related Experiment Video
Updated: May 4, 2026
11:16
Isolation of Protoplasts from Tissues of 14-day-old Seedlings of Arabidopsis thaliana
Published on: August 17, 2009
36.0K
Purification and viability determinations of plant protoplasts
1School of Botany, University of New South Wales, 2033, Kensington, N.S.W., Australia.
Planta
|January 17, 2014
Summary
A novel method purifies plant protoplasts using a specific density buffer, improving cell isolation. Fluorescein diacetate is also used to confirm the viability of these purified plant cells.
Area of Science:
- Plant biology
- Cell biology
- Biotechnology
Background:
- Plant protoplast isolation is crucial for genetic manipulation and cell fusion.
- Existing methods often yield low purity, contaminated with debris.
- Efficient purification methods are needed for successful downstream applications.
Purpose of the Study:
- To develop and describe an effective method for purifying plant protoplasts.
- To assess the viability of isolated protoplasts using a reliable assay.
Main Methods:
- A density buffer composed of 9.6% sodium metrizoate and 5.6% Ficoll was utilized.
- The method focuses on separating protoplasts from cellular and subcellular debris.
- Protoplast viability was assessed using fluorescein diacetate staining.
Main Results:
- The described method effectively purifies plant protoplasts from contaminants.
- High purity of isolated protoplasts was achieved.
- Fluorescein diacetate staining confirmed the viability of the purified protoplasts.
Conclusions:
- This density buffer method provides an efficient means of purifying plant protoplasts.
- The technique enhances the quality of isolated protoplasts for research.
- Fluorescein diacetate is a suitable stain for assessing protoplast viability post-purification.
