Zeiss Lightsheet Z.1, Руководство по эксплуатации

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CHAPTER 2 - SAMPLE PREPARATION
Carl Zeiss Specific Examples of Sample Preparation Lightsheet Z.1
32 000000-1790-528 02/2013
3.4 Preparation of a Plant Root ( Arabidopsis thaliana )
Plant research is an important field of investigation using plants as model systems. They are three-
dimensional objects that are difficult to image fully and are usually dissected and sliced before being
imaged and analyzed. This protocol has been used to mount complete young Arabidopsis thaliana plants
for imaging root development directly on the microscope.
Equipment and reagents
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1 % Low Melting Point (LMP) agarose in plant buffer
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1 ml syringes
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Arabidopsis thaliana seeds
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Heating block
Method:
1.
Several agarose beakers are prepared as described in the enclosed sample section.
2. Instead of pushing out the plunger to extract the agarose beaker, the plunger is pulled in to the
end of the syringe where it can be released leaving the beaker inside the syringe.
You need to make a long walled beaker to avoid inconvenient breakages and leakages that may be
caused by the following steps.
3. A seed of Arabidopsis thaliana is put at the bottom of the beaker.
4. The beakers are kept in the syringe in a humidified and well lit chamber to allow seed germination.
Fig. 18 Mounting an Arabidopsis thaliana root
(A) An Arabidopsis thaliana seed is positioned at the bottom of an agarose cylinder (B) After a few
days of development, the root can be seen in the agarose cylinder bottom. (C) The agarose cylinder is
pushed out of the syringe for imaging.