Non-Hardening Mountant Solutions
Glycerol-Based, High Refractive Index Solutions Reagents for Transmission Microscopy as well as Fluorescence Microscopy
All the following solutions have a refractive index of ~1.52 at room temperature (20°C) and are freely miscible with water. They may be used as an immersion oil in the standard way as well as a mountant solution. The refractive index of the materials makes them ideal for use when specimens are being examined by confocal laser scanning microscopy (CLSM)). They show very little absorption above 400nm.
The CFM range of mountant solutions should only be used with samples that have been fixed (4% paraformaldehyde in phosphate buffered saline for 30 minutes). The crosslinking (fixing) of the sample prevents non-covalently bonded fluorochromes, e.g. DAPI, Hoechst dyes, from becoming detached from the specimen etc.
If some bleeding of the dye(s) is still observed a further fixation with paraformaldehyde should be carried out. For sealing the coverslips, when CFM mountant solutions are used, the product CoverGrip® marketed by Bioutium has been recommended. (We are grateful to Dr. Sean Speese, Jungers Center for Neuroscience, Oregon,
U.S.A.for these many helpful suggestions).
The two images are examples of CitiFluor product applications, kindly provided by Dr. Sean Speese:
Above: 63x1.4 NA PlanApo/1 Airy/Nyquist sampling xyz/Zeiss LSM 710 Genetically expressed GFP-tagged nuclear protein in brain slice.
Below: 3D rendering of nuclei stained for LamDm0 in Drosophila tissue, mounted in CFM3 + antifade, imaged via confocal microscopy. Notice the staining intensity is consistent throughout the entire 55 µm stack with no increase in spherical aberration.
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