Visikol HISTO Beta Test Feedback - Light Sheet Microscopy
First, you should reach out to your imaging core manager and find out what type of light sheet microscope you have access to and what type of objectives it uses. Many light sheet microscopes such as the ZEISS Z.1 and the OpenSPIM will have water immersion objectives that are matched to a refractive index of 1.33. These objectives have two disadvantages for imaging tissues that have been cleared with Visikol HISTO-2: 1) The mismatch in refractive index between the objective and Visikol HISTO-2 (1.51) will limit imaging depth to approx. 3 mm due to spherical aberrations and overall sample thickness to 6 mm if you image from both sides, 2) The sample will need to be mounted in a cuvette that is placed within the imaging chamber.
The reason that the sample will need to be mounted in a separate cuvette is that the imaging chamber cannot be filled with Visikol HISTO-2 when using water immersion objectives as it is possible that the objective can be damaged by Visikol HISTO-2. Information on the preparation of samples cleared with solvent based techniques (Visikol HISTO-2, BABB, 3/i/uDISCO) for light sheet microscopy with water immersion objectives can be found here for the ZEISS Z.1.
However, if you have a solvent compatible light sheet imaging system like the LaVision BioTec Ultramicroscope then you can image directly in Visikol HISTO-2. The LaVision BioTec Ultramicroscope has been designed to work with almost every clearing solution (1.33 to 1.56) and will not get damaged by solvents.
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