Hi, I am trying to observe lignin through phloroglucinol staining with arabidopsis (petal and sepal).
The protocol I am using : 2% phloroglucinol in 100% EtOH , O/N
mount on slide
add HCl, coverslip
The problem here is that I keep getting the yellowish tissue. Lignin itself (at the end of the sepal and petal) is easily found with pink color, and other part (middle part of sepal and petal) supposed to look clear, but it is not.
I put them in 100% EtOH O/N, so I don't know whether it is caused by remaining chrolophyll or not.
Can somebody help me, please?
We haven't used this method much, but when we do, we usually put the tissue in Ethanol for at least a week before staining, changing the Ethanol at least once during that time. We also use 70% ethanol. Of course, because the plants we use are much thicker than Arabidopsis, we leave them in ethanol for a few weeks. This extracts pigments and other molecules that make your tissue not clear. It's also a convenient way to store them until you have time to do the staining. For Arabidopsis, you might try increasing the time that you leave the tissue in Ethanol prior to staining. I'd try 3 days as a start, changing the ethanol after 1 or 2 days. If that still doesn't look good, try a week. Good luck!
- Badges
- Science topic
- Similar topics
- Biological Science
- Biota
- Eukaryota
- Viridiplantae
- Streptophyta
- Plants