What is the best mounting medium to study plant cell on real-time?

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Molines Arthur

Hi,

Can you give more details please?

What do you want to do?

Do you want to mount a whole seedling, isolated plant cells from cell culture or a part of an organ like a portion of a leaf?

How long do you want to keep your sample under the microscope ? Is it for few minutes ? for hours ?

Regards,

Arthur

Joao Paulo Rodrigues Marques

Hi Arthur,

Thanks for your swift response.

As a plant anatomist, I am interested in study plant tissues. I will conduct some fresh sections and then I want to mount the slides to look for some modifications in plant cells/tissues. I don't know if this biological phenomenon occurs in minutes or hours, but it's better to make a medium that can stay for a long period. Is it possible? Should I use antioxidants agents?

Thanks for your time.

João Paulo

Hi João,

Arg, I have never done this...

Maybe the Hamant et al, Science paper in 2008 can inspire you. Check M&M of the Fig 4 they have grown apical meristem for minutes to hours in order to image them.

If you want to image the sample for a significant time you want to keep it "alive" so you need nutrients and sugar in the medium I guess. Antioxidants can help to avoid stress (maybe DTT).

If imaging lasts for hours I guees you will face two problems

1) the temperature will increase because of the laser (water will evaporate)

2) you will have to refresh the medium because nutrients and DTT will be consumed...

How to replace it ? No idea, maybe a chamber with a pump...

Sorry not to be able to help you more than that but it's out of my skills...

Good luck,

Thanks for your tips. I am thinking about it for some months, and I don't know how to do it. Well, if I use a widefield microscopy, do you think that it cause less damage than a confocal one?

Another important aspect, in your experiments, did you use PBS as a mounting medium? If my biological phenomenon occurs in some minutes, what is the best mounting medium?

Of course, that I might try every protocol... but it is the beginning...

Thank you for your help and time

João