Zeiss Lightsheet Z.1, Руководство по эксплуатации

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CHAPTER 2 - SAMPLE PREPARATION
Carl Zeiss Sample Mounting for LSFM Lightsheet Z.1
16 000000-1790-528 02/2013
The container can be easily molded using a gelling agent specifically chosen for its stiffness and
transparency. The custom-made molding system is made from a syringe where the plunger has been
modified to hold a cylinder of smaller diameter. This system allows tailoring of the size of the container
wall and is easy to use for molding (Fig. 11/
A ). The plunger is pulled into the syringe body and filled with
molten gelling agent. The plunger is further pulled to create the bottom part of the container
(Fig. 11/
B and C ). Alternatively, the system can be used to generate a hollow tube that can be
subsequently sealed (Fig. 11/
D
). The gelling agent is left to polymerize and then the tube is removed from
the modified plunger. The containers can be used directly or kept in a water-based buffer for later use.
The gelling agent must be transparent, and although the use of agarose is possible, the concentration
will depend on the size of the container walls and the inside chamber. It is recommended to use a higher
concentration of gelling agent to ensure the stability of the container. We have used a 1ml syringe as a
molding system and a concentration of 1.5 % agarose for the container molding. The stability is good
and the degradation of the optical path is minimal. Higher agarose concentrations may generate
aberrations.
Fig. 10 Enclosed chambers for LSFM.
Incubation chambers can be made by molding an agarose beaker that can be mounted on a simple
plastic holder and loaded with the sample prior to imaging (A). Another solution is to create a
chamber with a specific polymer with a refractive index close to water (for example PFTE or FPE) using
heat or glue to seal the chamber to the needed size and volume, and attach it to a suitable holder
(syringe, capillary etc…) (B). A PDMS, FPE tube or glass chamber can also be considered (C).
Fig. 11 Making and mounting an agarose incubation chamber.
A modified syringe plunger is made by inserting a smaller diameter cylinder on to the plunger (A, B).
The tube is molded by simply pouring the molten agent into this device (C). Once removed, the open
end of the container (D) can be closed with agarose (E).