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CHAPTER 2 - SAMPLE PREPARATION
Carl Zeiss
Sample Mounting for LSFM
Lightsheet Z.1
18
000000-1790-528
02/2013
2.3
Materials and Equipment
This section gives an overview on the materials and equipment for sample mounting and the sample
chambers of the Lightsheet Z.1. The generalization of the concept is also mentioned.
2.3.1
Sample Chambers
In the Lightsheet Z.1, the sample is positioned within a chamber containing an aqueous solution. This
chamber is tailored with O-rings to tightly fit the detection optics and avoid leakage. The upper part of
the chamber is open to allow introduction of the sample. The bottom can be equipped with a Peltier
Block or a Heatingblock (optional incubation). The remaining three sides are made in such a way that
glass coverslips can be fixed allowing entrance of the light sheets from two sides and observation of the
object by the user during the different steps of imaging using the appropriate software feature. The
original chamber is made of medical steel, however, depending on the buffer used (salt, pH etc.), the
experiment being performed (time lapse, live cell imaging etc.), there might be a need for more specific
chambers. Carl Zeiss provides the technical drawing of the sample chamber for Lightsheet Z.1 so that
users can develop their specific sample chamber
1
. You can refer to the sample chamber section of this
manual for further information.
When designing a chamber for your particular application you must take into account the following
points:
Transparency: user visual access, light sheet entry and exit routes, the distance of the coverslips for
the light sheet – and the water filled space in between - are a crucial measure in the optics
calculation of the Lightsheet Z.1 system. To ensure the functionality of the system these have to be
maintained when a custom made chamber is designed.
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Temperature control: heating devices, cooling devices
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Volume: size of the sample, buffer used (cost), drug treatment (cost)…
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Fitting: objective, illumination position, stage, heaters…
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Material: buffer, heater, sterilization, UV protection…
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Flow: flow entry and exit
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Size
−
Cost
1
Carl Zeiss Microscopy GmbH (hereinafter "we") hereby informs you that we will warrant the specified and agreed performance of the
Lightsheet Z.1 system only if sample chambers are applied and used that either are delivered or explicitly approved by us.
The sample chamber design has been optimized to ensure the most established applications of Light Sheet Fluorescence Microscopy. Exceptional
applications may require a slightly modified sample chamber design. In order to enable customized modifications of the existing sample chamber
we also provide the corresponding CAD file and a technical drawing. We explicitly advise you that already minor deviations of the dimensions
and tolerances specified in these documents will cause a significant loss of image quality and can potentially result in a liquid leakage. Therefore,
you will not hold us or one of our affiliates liable for any damages caused by the employment of self-built or third-party-built sample chambers,
the use of such self-built or third-party-built sample chambers will be solely on your own risk. Furthermore we want to inform you, that we will
not render any assistance relating to the production and application of such self-built or third-party-built sample chambers.
Содержание Lightsheet Z.1
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