STED Microscope


Figure 6: Unlocking gentle imaging in living cells. OMP25-SNAP expressed in HeLa cells labeled with abberior LIVE 610. Imaging in DMEMgfp-2 medium at room temperature. (A) RESCue imaging scheme consisting of three steps: confocal imaging, a pixel-wise decision to use or not use STED, and if the decision is made to use STED imaging. (B) Overview image taken before starting a time series in the marked area. (C) Every tenth frame of a time series spanning 50 frames. Scale bars: (A and C) 10 μm, (B) 20 μm.


volume or recording a time series. Tere are a range of deci- sion algorithms available to choose from, known as RESCue [15], DyMIN, and MINFIELD [16], depending on the require- ments and the type of sample. A special case is guided STED, a realization of adaptive illumination developed for malaria research [17]. It avoids exposing highly reflective iron deposits that appear in late stages of the parasite’s life cycle. Normally,


2022 July • www.microscopy-today.com


these deposits limit imageability and cause phototoxicity, but avoiding these sensitive areas allows non-invasive imaging of these difficult samples. Aberration-corrected deep tissue imaging with 3D-super-resolution microscopy. Tissue imaging,


like


live-cell imaging, is a challenging application, not only because sample preparation is more difficult, but also from


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