1. **Fixation**: Tissues are fixed using a formalin solution³. This step preserves the structure of the cells and tissues in the sample.
2. **Trimming and Transferring**: Fixed specimens are trimmed to the appropriate size and transferred to labeled cassettes³.
3. **Dehydration, Clearing, and Embedding**: The samples are then dehydrated, cleared, and embedded in paraffin wax³. Dehydration involves removing water from the sample, usually with alcohol. Clearing makes the sample transparent and prepares it for embedding.
4. **Sectioning**: Sectioning with a microtome produces thin tissue slices which are transferred to glass slides³.
This is a general protocol and might need to be adjusted depending on the specific type of plant seed you are studying. For example, protocols have been developed for rapid clearing and staining of fixed Arabidopsis ovules¹, and for studying crassinucellate ovules of the sugar beet².
Please note that these steps should be carried out in a lab under the guidance of a trained professional, as they involve the use of chemicals and specialized equipment. Always follow safety guidelines when handling chemicals and equipment. If you're planning to do this in a lab, make sure you have the necessary permissions and guidance.
(1) Histology Slide Preparation: 5 Important Steps - Bitesize Bio. https://bitesizebio.com/13398/how-histology-slides-are-prepared/.
(2) Protocol for rapid clearing and staining of fixed ... - Plant Methods. https://plantmethods.biomedcentral.com/articles/10.1186/s13007-019-0505-x.
(3) Refinement of a clearing protocol to study ... - Plant Methods. https://plantmethods.biomedcentral.com/articles/10.1186/s13007-019-0452-6.
(4) An optimized histological proceeding to study the ... - Plant Methods. https://plantmethods.biomedcentral.com/articles/10.1186/s13007-020-00604-6.
