Methods used in the lab
Structured Illumination
a linear grid is inserted into the field stop plane and thereby imaged into the object plane. By taking three images with the grid being shifted by Δ=2/3Φ, all parts from the sample are illuminated at least once. An optically sectioned image is extracted mathematically.
Comparison of conventional epifluorescence (left) and grid-SIM (right) in the OB.
The essential feature of the confocal laser scanning microscope (cLSM) is the generation of optical sections by removal of out-of-focus light. About ten years ago structured illumination microscopy (SIM) was introduced as an alternative method of obtaining optical sections from biological specimens.
We compared the resolution performance of the commercial SIM-system (ApoTome) to a commercial cLSM-sytem (510, both Zeiss, Jena, Germany) (Weigel et al., 2009). We found that, depending on the particular conditions, the ApoTome achieves a similar resolution as the cLSM. For imaging deep inside tissue however, the cLSM is preferred. We use grid-structured illumination for the imaging of multiply labelled cell cultures and histological slices.