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Protoplast Iolation Experiment: Enzymatic method

At LJMU Byrom Street lab, together, me and Dr Symonds looked at protoplast isolation, which is an experiment that breaks down the cell wall to reveal a single cell.


This experiment was first exhibited in 1960, Cocking used a concentrated solution of cellulase enzyme, prepared from cultures of the fungus, Myrothecium verrucaria, to degrade the cell walls (Protoplast isolation, 2020).


During a meeting Dr Symonds suggested the experiment and we both agreed it would be interesting to conduct and photograph. The image could then be shown in the infographic zine which will be used as part of Mindful Garden: Visualising the Unseen project. This will help children visualise what a single cell looks like without a cell wall. Likewise the experiment bridges art and science together to demonstrate the important link of art-science used in education.


Image 01: Showing enzyme fluid and plant specimens.


Image 02: Showing single cell which could be seen under the microscope.


To conduct the experiment leaf tissue is preferred as it allows the isolation of a large number of uniform cells. The activity of the enzyme depends on pH, however, it is recommended to be between 4.7 and 6.0. Generally the temperature of 25–30°C is sufficient for isolation of protoplasts. The duration of enzyme can be as short as 30 minutes or as long as 20 hours. In this experiment it was only for 30 minutes.


Reference

Protoplast isolation. (2020). Available at: Cocking used a concentrated solution of cellulase enzyme, prepared from cultures of the fungus, Myrothecium verrucaria, to degrade the cell walls.

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