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BP: Fachverband Biologische Physik

BP 20: Poster VIII

BP 20.24: Poster

Dienstag, 17. März 2020, 14:00–16:00, P2/4OG

Point Spread Function engineering in real time for iSCAT — •Vivien WALTER and Mark WALLACE — Department of Chemistry, King's College London, London, United Kingdom

Interferometric scattering (iSCAT) microscopy extends the limits of traditional optical microscopy by imaging the interference pattern between light scattered by an object and a reference beam reflected from a surface. Efficient subtraction of the large reference signal and its magnitude relative to the scattering of the object is key to high-speed high-sensitivity imaging. However, subtraction of the reference signal in complex biological samples requires more sophisticated processing to detect single biomolecules.

Optical processing of images is commonly used in microscopy to optimise the collected signals by increasing contrast or decreasing noise. Fourier plane processing is a common method to select specific image properties and while powerful the application of optical processing to iSCAT has typically been limited to static apodising filters. Adaptive Fourier filtering can be performed using Spatial Light Modulators (SLM), computer controlled high resolution devices capable of applying any type of filter in real-time.

We demonstrate here the application of real time SLM processing applied in iSCAT microscopy to obtain a 6-fold increase of the signal-to-noise ratio. We investigate a range of optical processing methods, and demonstrate that this optical pre-processing reduces the molecular weight threshold of detectable label-free proteins and polymers.