Zeiss Lightsheet Z.1, Operating Manual

Page: 67 / 427

CHAPTER 2 - SAMPLE PREPARATION
Lightsheet Z.1 Sample Mounting for LSFM Carl Zeiss
02/2013 000000-1790-528 11
In the case of Lightsheet Z.1, the Sample Starter Kit provides four types of color coded capillaries and a
specific sample mounting device with color coded sleeves to fit each type of capillary perfectly to the
sample holder. Moreover, a syringe sample holder is provided.
Below the preparation of a syringe for large sample and a basic protocol for capillary mounting are
discussed.
An example of how to tailor a syringe for Lightsheet Z.1 sample preparation is illustrated in Fig. 5. The tip
of the syringe is cut off to create an even cylinder, and the gel solution is pumped in using the plunger.
The sample is then positioned precisely within the gel.
After the gel has polymerized, the plunger is used to push out the specimen prior to imaging.
Imaging is not done through the syringe or capillary, since this would impair the image quality due
to the optical properties of the material.
For smaller samples, a capillary can be used as a sample embedding container. There are several
commercial companies that provide glass capillaries with specific Teflon plunger. The Lightsheet Z.1
sample preparation kit comes with four sizes of capillaries and their specific plunger for this purpose.
Make sure you use the right capillary for your sample. The sample size should not be more than 2/3
of the final agarose diameter and no less than 1/3. You should also ensure that you use the right
plunger for you particular capillary. Finally, the Teflon plunger should be handled carefully and
checked regularly for integrity to avoid leaks that will lead to sliding of the gel rod.
Fig. 5 Preparing a sample embedding container with a syringe.
Many tubular objects can be used to make embedded samples. A simple technique is illustrated here.
The tip of a syringe is cut away (A, B), agarose can then be easily pumped in using the plunger (C), and
the sample can be positioned within the agarose tube (D). After polymerization the sample can be
pushed out of the syringe for imaging (E).