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Rolly Wiegand – CALM Leica SP5 manual
3
Important notes regarding the use of objective lenses:
It is vital for the longevity of the lenses that the correct immersion medium is used. Please
double check which lens is in place before you apply any immersion fluid. If you
accidentally made a mess, do not try to cover it up, but report it immediately to the CALM
staff.
You must clean your lens and your sample before switching to a different immersion
medium.
In case you would like to use the 63x WATER NA 1.2 lens, which is usually not on the
turret, please let the facility manager know in advance. This lens has a collar for adjusting
the lens to correct for different coverslip thickness. The black mark indicates the correction
at room temperature, whereas the red mark indicates the corrective settings for 37
°
C.
Fig. 4
Choose the lens you want to use by matching the refractive index (RI) of the immersion
medium with the medium embedding your sample (oil lens with a fixed sample, mounted in
a high RI medium; water lens with a live specimen in aqueous solution). Mismatch of the
RIs within your ‘optical sandwich’ can cause unwanted internal reflection and blurring. Also
bear in mind that the higher the NA of the lens, the higher the resolving power. If you have
the choice between two lenses with different magnification factors but the same NA (e.g.
63x or 100x oil, NA 1.4), the actual magnification should be adjusted with the system zoom
(see 4.7.4.) and should fulfil the Nyquist sampling criterion. This means the two lenses will
give you the same magnification and resolution, but the lens with the lower magnification
factor will allow you to image a larger field.
2. Operation of the DMI6000 inverted microscope base
2.1. The microscope should be already switched on via the TCS control panel (see above
section 1.2.).
