Please make sure that the coverslip is centered on the glass slide in order to fit into the sample holder.
See below pictures of the ZEISS level adjustable piezo stage insert.
Fix your cells according to your standard protocol. For SR-SIM imaging, a thoroughly clean glass surface plays a crucial
role. Therefore it is beneficial to seal or attach the coverslip in a way that facilitates cleaning with ethanol, without
moving the coverslip.
Fluorescent Labels for SR-SIM
All common types of organic dyes usually conjugated to antibodies or fluorescent proteins are suitable to be used for
SR-SIM. Make sure to have a highly specific labeling with low background to obtain a good signal to noise ratio.
For multicolor samples the fluorophores should be selected for minimal spectral overlap to avoid crosstalk. Available
filter sets for ELYRA PS.1 are are optimized for the available laser lines: 405, 488, 561 and 642 nm.
Note
Cytosolic or other non-specific fluorescent protein expression (e.g. GFP) will result in staining of extended areas.
Since well-defined structures are missing to interfere with the grid pattern, modulation contrast will be low and the
final image will lack high resolution information.
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Figure 1
ZEISS level adjustable piezo stage insert
