Zeiss Lightsheet Z.1, Руководство по эксплуатации

Страница: 83 / 427

CHAPTER 2 - SAMPLE PREPARATION
Lightsheet Z.1 Sample Mounting for LSFM Carl Zeiss
02/2013 000000-1790-528 27
As in LSFM there is no need for oil or any specific chemical for imaging, the cleaning of samples is not
necessary. However, you can rinse the sample within the capillary or syringe with water or your specific
buffer after imaging if the chamber was containing particles, bacteria or other chemicals (dyes, drugs
etcetera).
Labeling the samples can be an issue as it can be tricky to mark the name of every sample on the
embedding container (capillary, syringe…). A simple marking technique is to use tape roll around the
syringe plunger or the capillary. This must not affect the handling of the sample on the microscope.
Another approach is to number the sample and to register the detail on a lab book. However, this can be
a problem if you store many sets of samples in the same fridge day after day.
Sample preparation techniques usually allow long time storage (paraffin embedding, slides…). As long as
a few basic rules are followed (keeping away from light, temperature…) they can be kept up to years. In
the case of LSFM, the samples are imaged in a water environment and must be always kept wet, even for
long time storage. This can be a challenge. Usually, we keep fixed samples in the fridge using a sample
embedding container support and we refill the buffer tank from time to time. However, we never kept
samples for more than a month under such conditions. A longer storage possibility is to use a water tight
container where the samples are kept with enough water not to dry out. One point to consider is the
way the sample was prepared. Embedded samples may weaken with time as some gels may not maintain
their strength over time at 4 °C. Hooked samples may as well be loosening and fall from their support. It
may be better to unhook them and store them in a different type of container.
Fig. 15 Supports for sample embedding containers.
Support are used to hold three dimensional objects that cannot be held flat easily. Moreover,
embedded samples need to be kept in buffer to avoid gel shrinkage or sample damage. A simple
system is to use a beaker filled with PBS and place plasticine on the upper border to support the
sample embedding container (A). More elaborate supports can be made using clips of different sizes
for holding syringes (B) or even capillaries (C).