Zeiss Lightsheet Z.1, Руководство по эксплуатации

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CHAPTER 2 - SAMPLE PREPARATION
Carl Zeiss Sample Mounting for LSFM Lightsheet Z.1
8 000000-1790-528 02/2013
2.2 Holding the Sample
In LSFM, the detection axis is at 90° from the illumination axis. There are two main approaches to design
such an optical configuration: horizontal or vertical, with respect to the detection axis. In both cases, the
sample must be positioned at the intersection between the two axes in order to be observed. The
Lightsheet Z.1 is a horizontal LSFM implementation and so the sample is presented from above, hanging
along the gravitation axis to be scanned through the light sheet in order to acquire stacks of optical
section images. Several possibilities exist to hold the sample in such an optical configuration.
In a vertical configuration, the simplest way is to place the sample on a slide or a cuvette filled with
medium underneath the objective (Dodt et al., 2007), alternatively the sample can be embedded in a gel
rod that can be rotated. In a horizontal configuration, like the Lightsheet Z.1, the sample can be either
embedded in a stiff gel (Fig. 3/
A ) (Huisken et al., 2004) hooked and positioned in front of the objective
(Fig. 3/
B
) (Engelbrecht et al., 2007), placed in a container (Fig. 3/
C
) (Engelbrecht et al., 2007, Kaufmann
et al., 2012, Pampaloni et al., 2007) or placed on a slide and positioned at a 45° angle (Fig. 3/ D ).
Alternatively, some investigators are using a system presenting the sample from underneath for better
stability (Huber et al., 2001).
The Lightsheet Z.1 is not designed to support mounting on a coverslip (Fig. 3/ D ).
Fig. 2 Different sample types.
(A) very large objects (cm), (B) large objects (mm), (C) medium size samples (100 µm) and small
samples (10 µm) (D). Each is represented in relation to the following one to allow size comparison.