Hello! We want to stain in our lab samples of Sempervivum infected by Endophyllum sempervivi. We would appreciate any suggestion of a cheap, quick and effective staining method to observe hyphae or other structures within the leaves. We just want to screen different parts of the plant to see if the fungus is present before showing any disease symptom, so a staining technique that allows us to stain the whole leaf would be also acceptable.
Thank you in advance!
Hello, there are several methods to dye fungi in tissues, whether they are vegetable or animal. Before I can help you, I need to know what fixation treatment you are giving to your tissue.
There are direct methods such as Wright and Wright Giemsa staining and indirect methods such as cotton blue or Grocott staining but to know which one to recommend and how to apply it I need to know what type of fixation you are using and if you give a previous treatment to weaken the cell wall of the tissue
Thank you very much for your response. We are quite unexperienced in this area. Our only previous experience is with the staining of arbuscular mycorrhiza with the ink and vinegar technique. We tried to clarify leaf samples using a similar method to that of roots (KOH 10%) but it did not work at all. We've never done anything similar in leaves, so we do not really know what kind of fixation treatment we should apply to the leaf samples. We'd appreciate your feedback.
To stain pathogenic fungal structures within plant leaves, you can use staining techniques that specifically highlight the fungal structures. Here are some common staining options you can consider:
Stains with fungal-specific dyes: There are a variety of fungal-specific dyes available that can selectively stain fungal structures within plant leaves. Some examples are lactophenol blue staining or lactophenol-cotton blue stains, which stain fungal structures blue while plant cells remain colorless.
Aniline Blue Stains: Aniline blue staining (Lactophenol Aniline Blue Stain) is a widely used technique for staining fungal pathogens in plant tissues. This stain highlights the fungal structures in dark blue or purple, allowing good visualization of fungi on plant leaves.
India ink stains: India ink staining is a simple technique that can be used to highlight fungal structures on plant leaves. It consists of immersing the sample in diluted India ink and then rinsing it to remove excess ink. This stains the fungal structures, while the plant cells remain unstained or lightly stained.
Fluorescent Stains: If you want to visualize fungal structures with higher contrast and sensitivity, you can use fungal-specific fluorescent stains. These stains contain fluorescent dyes that bind to fungal structures and fluoresce when excited with ultraviolet light. Examples of fluorescent stains for fungi are Calcofluor White and DAPI (4',6-diamidino-2-phenylindole).
Thank you very much! We'll try some of those methods. If anyone else has another suggestion, we'll be glad to listen.
- Badges
- Science topic
- Similar topics
- Analytical Chemistry
- Extraction