Reference no: EM132787547

Background Information

Rhonda was doing a project on the fungal pathogen of barley, Ustilago hordei. When she began her work, the prevailing belief was that U.hordei was only pathogenic after it had mated and formed a dikaryon, a cell with to haploid nuclei. The question she try to answer was " Can a monokaryon having one diploid nucleus initiate infection as well?" Her task is to create diploid monokaryons in the lab. However, she was not certain if the procedure she planned to use would give the desired result. The only way she would know for sure as to look at the cells after completing the procedure and determine if they contained one or two nuclei. The nuclei of U. hordei are not easily seen in a wet mount, so Rhonda needed to stain them. She began by trying DAPI, a fluorescent dye that binds DNA. After staining cells with DAPI, she could see nuclei clearly, but she had forgotten that the rest of he cell would not fluoresce, it was difficult to determine if two fluorescing bodies close to each other were the two nuclei of a dikaryon, or the two nuclei of overlaping or adjacent monokaryotic cells. She used another dye, Calcofluor, to specifically stain the cell walls. The two dyes together solved her problem. She could easily distinguish dikaryons from monokaryons. Rhonda's experiments showed that diploids are also pathogenic

Question

-There is a nonfluorescent dye that specifically stains chitin in fungal cell walls that you may have used- LBCP -Would double-staining cells with that dye and DAPI have solved Rhonda's problem? Explain

-List three other fluorescent stains. What is the usefulness of each?

-Describe how simple staining and fluorescence staining are similar and how they are different. What are the advantages of each?