Dear Experts!
I have compared the stomatal features of leaves from Musa species. Mechanical peeling opted from fresh as well as preserved leaves (fixed in FAA - 1:1:3 ratio for 24 hr). I found significant differences in the size/ diameter of the stomata. Will leaf preservation in FAA alter the size of stomatal apparatus?
Hi! So, the fixation of the leaf in FAA shouldn't alter the stomata size, since this solution interrupts any biochemical reactions that may be occuring at the moment and preserves the tissue aspects. I normally use PFA 4% (a solution of paraformaldehyde, that is basically a polymer of formaldehyde), and never had any problem with it - but is important to pay attention in the volume of the samples and the volume of FAA that you are using, plus the specific time necessary to the fixation of the tissue to be done successfully (with PFA 4% I usually leave the samples for 48-72h in the fridge and in sequence dehydrate them in alcoholic series 10-70% for storage, also in the fridge).
FAA fixation can save from alteration in stomata size, but care may be taken during collection of samples from the field. Time lag in preservation causes problem in tropical climate zone
Dear M. Manokari,thank you for share the result.Previously I have tested the mehtod to observe size and shape of the stoma, no abnormal were found after the treatment of FAA.
Dear Experts Mayara Alves Pereira, Dr Justin R Nayagam & Huiling Ma
Thank you very much for your reply, I have used the recommended concentration of FAA, which I am using for 8 years in almost 30 plants. So far, I've never noticed that changes in stomatal morphometry other than Musa. Let me check it on other Monocots. Thanks once again for your valuable time.
-Regards
It will not alter the size of the stomata, as it is already differentiated cell, and also with a cellulosic cell wall, which maintains cell shape and size, as cellulose is a very strong polymer. Hence, the size of stomata will not alter during FAA fixation. However, if you compare Carnoy's fluid, FAA, Gluteraldehyde with respect to organelles and nuclear material preservation. Carnoy's fluid is best preservative over FAA, while Gluteraldehyde is best over all other preservatives and used for ultra structural studies.
I think stomata size means here the size of stomatal pore which used to open and close based on the surroundings (air or chemicals). Stomata may close at some extent while fixing the leaves or peel in fixative because the fixative may enter in the cells by passive diffusion.
Well said Mahipal Shekhawat Sir,
Changes in stomatal pore size were observed before and after fixation. Thank you.
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