My group mates and I were tasked to prepare "perfect and permanent" slides for one of our classes. Sadly, it did not work out the way we wanted it to. As a result, we created some over-stained and some under-stained samples and slides that had bubbles.
When placed under a microscope, we found some black structures that were inside the cells. We want to determine what they are, since there aren't that many studies about the Ficus nota. It may be a component of the cell, a contaminant of some sort, or dirt from the slide.
We used safranin as the stain. Through research, we have not found any sources that say anything about safranin staining creating black components, other than the cell wall, given the right procedures. The sample preparation procedure involved the leaf undergoing killing and fixing solution (made up of ethanol, formalin, and acetic acid), bleaching solution (bleach, NaOH, and water), various ethanol and xylol series of increasing concentrations, and staining by safranin.
It would be much appreciated if one of you could help us with our project. If there are any further questions about the study, feel free to ask. But bear with me due to my limited knowledge on the topic, as I am still a student. Thank you!
Dear Danielle
The best way to answer your question is to follow scientific path. you have a question so we could have some presumptions to reach the right answer.
Safranin is a biological stain used in histology and cytology. Safranin is used as a counterstain in some staining protocols, colouring all cell nuclei red. This is the classic counterstain in both Gram stains, and endospore staining. It can also be used for the detection of cartilage, mucin and mast cell granules.
According to previous information you should ask some primary questions of yourself and try to answer them then you can have the answer.
1- What was the aim of this staining?
Probably, staining the ficus nota cell nuclei
2- have you reached that goal?
Yes
3- What else could be happened during staining, which has affected the results?
- Non-living contaminations on samples (use fresh, healthy, rinsed leaves)
- Non-living contaminations on glass (use Disposable slides)
- Presence of Inorganic matter inside or outside of cells (ask if your plant accumulate such coumponds)
- Contamination of samples with bacteria (safranin could stain bacteria too so make sure that plants are not infected or contaminated)
- It is unlikely the presence of cartilage or mucin there but you can consider this too.
If the presented offers didn’t work you might have found something new.
I wish the best for you
Hello Danielle,
I think it came probably from yours staining solution. what concentration you used for staining, what was the concentration of solution prepared, and what kind of sample you have utilized for staining and when did you picked up for visualization under microscope? what I may suggest you, pic a small drop of only staining solution on slide and first observe this. Make fresh solution properly for staining and visualized a fresh plant sample with it. when you observe those black dots will help you finding what is it.
good luck
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