Problem: water immersion lens on inverted microscope is a problem because 1.) water breaks surface tension and runs down objective, 2.) water evaporates, 3.) Immersol W or other similar immersion media not thick enough to hold contact with long working distance objectives.

Solution: Use something with a refractive index matching water that is thick and goopy and resilient to evaportation. GenTeal was recommended on the confocal microscopy listserv.

This webpage shows how to use GenTeal for microscopy and shows a few of our first images taken with this stuff. In addition to images shown here, we have imaged hair follicles in a live mouse with the incubator chamber on for more than 4 hours without evaporation, other loss of contact, or needing to add more GenTeal.

This shows a lymph node being excited for imaging at 920 nm.

Click on this image to see a big version half the original resolution of a single optical section collected by multiphoton microscopy with the Zeiss 20X N.A. 1.0 water immersion objective pictured above.
It shows a mouse nail with macrophages in yellow (CD11c-YFP), second harmonics of collagen in blue, and keratin or other background in green.

Click here for a 3D movie collected by multiphoton.
Mouse footpad same labels as nail above.

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