Embedding
Ideally, the sample should be embedded in a medium that matches the refractive index (RI) of the immersion oil
(n=1,518). The following media perform well, despite having a lower refractive index.
• Fluoromout-G (SouthernBiotech) with RI of 1,40.
• 86% Glycerol with 2.5g/l DABCO (1,4-deazabicyclo[2.2.2] octane, Sigma Cat. D2522) in Tris-HCl. 1M pH 8.0.
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Sample requirements
• Non-hardening VECTASHIELD Mounting Medium with RI of 1,44.
• ProLong Gold (Life Technologies). Be aware this embedding medium needs 2 day to harden (in order to reach a
constant RI). In addition your sample may shrink during this process. Refractive index (RI) of the cured product:
1,46.
• SlowFade Gold (Life Technologies) with RI of 1,42.
• 2,2’-thiodiethanol (TDE) – aqueous. RI can be varied ranging from being that of water (1.33) to that of immersion
oil (1.52) by appropriately diluting with water.
Note
In order to have stable imaging conditions, especially concerning the RI of the mounting medium we would urge
you to prepare the slides at least one week before use. Please note, that during curing the RI will rise.
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