Hi all,
I am studying root responses to gradient treatments on Arabidopsis roots using a confocal microscope. Treating roots is fine but when you place them on the slide you no longer know which side was closest to the treatment.
How can I perform confocal observations still knowing what side of the root is which?
I was thinking about growing the roots between the slide and cover slip but I am not sure if this is doable.
Has anyone done something similar?
Thanks in advance.
Hi, Daniel,+yes, you can growth slides between slides with spacer, but you can alos very carefully take plants from agar, carefully stain it (without changes root position) and carefully mount on the slides.
This is very easy if ypou use FP markers and do not want to perform additional staing. In this case you can easy recognise root side even on the slides.
I dont know if it would work with your setup, but you may be able to look at them in situ with an immersion lens. I've done this to look at very delicate tissues in small petri plates.
Might it be possible to stain the roots while intact on the plant and with a little bit of agar? When you are ready to put them on then the sliede, THEN you cut, even on the slide...
I assume you cut the roots when you make your prep. If I were going to do this I would use the very simple method of cutting the root at an angle, so that the angled surface always bears a predetermined relationship to the treated side. That way, as long as you can see the cut end you will know which side has been treated.
Thanks for the suggestions everyone. I am going to try fixing the root/plant with water on the slide and place a cover slip on it. Then I intend to add the treatment from one side with a pipette and suck up the water from the other side with some paper just a bit so the treatment goes inside and then diffuse towards the other side of the slide. I will let you know if it works.
- Badges
- Science method