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I agree, for live cell imaging water immersion lenses on inverted
scopes with cover-slip bottom chambers (Matek and others) is best, in my
opinion. For live mice it is much easier to put them on the inverted
scope. The Zeiss LD C-Apohcrmat 40x 1.1NA water lens is terrific, I do
agree 40x water is great, but if you do need the higher mag the Olympus
60x 1.2 NA Uplanapo is a terrific lens. Zeiss Immersol W is highly
recommended, because, in my lab, at least, due to the dry conditions in
the winter immersion water evaporates away very quickly and Immersol W
will be better for long-time imaging than going in and adding more water
every hour. We image up to a few days at physiological conditions so
focus correction is critical, also. Focus correction is very serious if
you have confluent or non-confluent single layers of cells - and
physical techniques (Nikon Perfect Focus with internal laser diode and
total internal reflection) are better than image-based focus correction
techniques, which need a lot of finessing and can zoom in on the
brightest feature, such as dirt. 40x and 60x water lens are usually
the most carefully designed and best for the widest possible variety of
experiments - fluorescence, TIRF, confocal, 2-photon, etc. if you have
the money.
Best,
Evangelos Gatzogiannis
Advanced Biological Imaging Scientist
Harvard University Center for Nanoscale Systems
11 Oxford Street
Cambridge, MA 02138
617-496-8250
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