1. PicoSIM – The idea | ||||||||||
As a wide-field technique, SIM is potentially significantly faster than scanning techniques such as confocal microscopy. However, the requirement to acquire three images reduces its speed. Polarised illumination coded Structured Illumination Microscopy (PicoSIM) encodes the three individual light patterns needed for conventional SIM in the polarisation of the illumination light.Benefit: Acquisition of complete SIM data in one single exposure. This would allow ultrafast optical sectioning with high spatial resolution in all three dimensions. |
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2. How it works |
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Illumination
Effective Illumination
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3. Performance |
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Because of fluorescence anisotropy the full degree of input polarisation cannot be preserved. Hence, picoSIM images recorded after polarisation filtering correspond to conventional SIM images with reduced illumination pattern contrast. This leads to a reduced signal-to-noise ratio as compared to conventional SIM. | ||||||||||
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4. Setup |
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Illumination
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Detection
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5. Preliminary results |
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Data acquired using coherent laser illumination and an additional magnification on the illumination side of the system. Furthermore, just one analyser instead of three was used on the detection side, i.e. the emission light was not yet split into 3 components, but the analyser was successively rotated. (Objective: 100x, 1.4 NA oil immersion) |
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