Fungal-modified hydrogel-discs
Creating fungal-modified hydrogel discs involves several steps, each critical for ensuring the quality and functionality of the final product. This protocol outlines a general approach to producing these discs, incorporating fungal treatment to modify the properties of hydrogel. The specific details may vary based on the type of hydrogel and fungus used, as well as the intended application of the product.
Materials Needed:
Hydrogel precursor (e.g., agarose, polyacrylamide)
Fungal culture (type depends on the desired modification)
Culture medium for fungi
Petri dishes or similar containers
Sterile water (if necessary)
Standard laboratory equipment (incubator, laminar flow hood, autoclave, etc.)
Protocol:
Preparing the Hydrogel:
Hydrogel Solution Preparation:
Dissolve the hydrogel precursor in sterile water or an appropriate solvent to create a solution at the desired concentration.
Heat the solution if necessary (e.g., agarose) to ensure the precursor is fully dissolved.
Sterilisation:
Sterilise the hydrogel solution using an autoclave or by filtration to prevent contamination.
Casting Hydrogel Discs:
Pour the sterile hydrogel solution into molds of the desired shape and size.
Allow the hydrogel to cool and solidify at room temperature or in a refrigerator.
Fungal Culture Preparation:
Inoculum Preparation:
Revive the fungal culture from a stock or obtain a fresh culture.
Grow the fungus in a suitable culture medium until it reaches the desired growth phase.
Harvesting Fungal Cells:
Harvest fungal cells or mycelium by filtration or centrifugation.
Wash the collected fungal material with sterile water to remove residual media.
Hydrogel Modification:
Inoculating Hydrogel with Fungus:
Place the hydrogel discs in a sterile container, such as a Petri dish.
Inoculate the hydrogel discs with the fungal cells or mycelium, ensuring even distribution.
Incubation:
Incubate the inoculated hydrogel discs at optimal conditions for fungal growth (temperature, humidity, and light).
Monitor the growth regularly and maintain sterile conditions to prevent contamination.
Harvesting and Post-treatment:
Once the desired level of modification is achieved (e.g., enzymatic degradation, polymerization), terminate the fungal growth by changing the conditions (e.g., temperature shift, drying).
Wash the hydrogel discs to remove any non-adhered fungal material.
Characterisation:
Characterise the modified hydrogel discs using suitable methods to assess the extent and nature of modification (e.g., mechanical testing, microscopy, chemical analysis).
Storage:
Store the fungal-modified hydrogel discs under appropriate conditions until use. Ensure that the storage conditions maintain the stability and functionality of both the hydrogel and the fungal modifications.