flow cytometry

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greenfish
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flow cytometry

for classroom teaching what is the best protocol to follow for estimation of nuclear dna in plants using flow cytometry and microdensitometry?
can one use dehydrated tissue instead of fresh ones? doable answers will be welcome

Ivan Delgado
Ivan Delgado's picture
 

 
Hi greenfish,
Are you trying to use flow cytometry to analyze plant nuclear dna? I ask because it does not seem to be the best approach for either application. In other words, if you want to demonstrate flow cytometry to a class I would recommend using some of the standard kits available that are based on different-sized beads. They work great and you do not need to go through the hassle of dealing with cells. On the other end if your main objective is to work with plant cells, it would be quite tricky to analyze them by flow cytometry unless you are using plant cell cultures and generate protoplasts. 
If you provide more details maybe I will be able to be more helpful. I've never analyzed dehydrated cells by flow cytometry, and my guess is that it would not work very well.

greenfish
greenfish's picture
thanks ivan for the inputs.

thanks ivan for the inputs. well i am going to try some expts and dont know how to start. u can call me a beginner so i have to prepare best for class. theory of the flow cytometer i already have ready with me. so now i have to get the practical demostration.

have you used this technique?can you help me as to the best way to do it? i did not know that we can do it with kits. whcih company kits are these?
as i am a novice in this technique thats why i asked the forum members fpr suggestions.

hoping to read your suggestions soon.
best wishes

Ivan Delgado
Ivan Delgado's picture
 

 
Hi greenfish,
No problem. Yes, I have used this technique in the past but I currently do not use it. What I would do if I were you is ask the vendor that sold you the flow cytometer for a kit that best demonstrates the capabilities of the instrument. All vendors should have one for that purpose. Trying to use other kits would likely work, but would require  you to do extra work figuring out how to adapt the kit to your flow cytometer. 
Good luck