Abstract
We demonstrate the reconstruction of images obtained by multifocal structured illumination microscopy, MSIM, using a joint Richardson–Lucy, jRL-MSIM, deconvolution algorithm, which is based on an underlying widefield image-formation model. The method is efficient in the suppression of out-of-focus light and greatly improves image contrast and resolution. Furthermore, it is particularly well suited for the processing of noise corrupted data. The principle is verified on simulated as well as experimental data and a comparison of the jRL-MSIM approach with the standard reconstruction procedure, which is based on image scanning microscopy, ISM, is made. Our algorithm is efficient and freely available in a user friendly software package.
Highlights
Optical super-resolution, the resolution of microscopic details below the diffraction limit, is revolutionising biomedical research
We demonstrate the reconstruction of images obtained by multifocal structured illumination microscopy, MSIM, using a joint Richardson–Lucy, jRL-MSIM, deconvolution algorithm, which is based on an underlying widefield image-formation model
MSIM is the parallelised version of image scanning microscopy, ISM [12], which uses the inherent super-resolving potential of confocal microscopy to double image resolution, when the detection pinhole is nearly closed
Summary
Optical super-resolution, the resolution of microscopic details below the diffraction limit, is revolutionising biomedical research. A particular modality of structured illumination microscopy is called multifocal SIM, MSIM [11] In this variant multiple diffraction limited spots, rather than sinusoidal illumination patterns, are used to achieve super-resolution and the pixels on a widefield detector act as tiny pinholes, akin to a parallelised version of confocal microscopy. MSIM is the parallelised version of image scanning (confocal) microscopy, ISM [12], which uses the inherent super-resolving potential of confocal microscopy to double image resolution, when the detection pinhole is nearly closed [13]. For these reasons, the conventional algorithm for MSIM reconstruction [11] uses a confocal image formation model: i( x )⃗ = s( x )⃗ ⊗[e( x )⃗ · PSFdet ( x )⃗ ],
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