In this work we have explored the live-cell friendly nanoscopy method Multiple Signal Classification Algorithm (MUSICAL) for multi-colour imaging of various organelles and sub-cellular structures in the cardiomyoblast cell line H2c9. We have tested MUSICAL for fast (up to 230Hz), multi-colour time-lapse sequences of various sub-cellular structures (mitochondria, endoplasmic reticulum, microtubules, ...

Microscopy is possibly the best tool we have to peer into the microscopic world to enhance our understanding of the usually invisible, but highly complex and vital events every moment taking place inside living cells. Microscopy is brilliant, but also has its physical constraints and technical limitations. Technical advances have in the last decade pushed optical microscopy past physical limits ...

Mitochondria are susceptible to damage resulting from their activity as energy providers. Damaged mitochondria can cause harm to the cell and thus mitochondria are subjected to elaborate quality-control mechanisms including elimination via lysosomal degradation in a process termed mitophagy. Basal mitophagy is a house-keeping mechanism fine-tuning the number of mitochondria according to the metabolic ...

In 1934, Frits Zernike demonstrated that it is possible to exploit the sample’s refractive index to obtain superior contrast images of biological cells. The refractive index contrast of a cell surrounded by media yields a change in the phase and intensity of the transmitted light wave. This change can be due to either scattering or absorption caused by the sample. Most cells are transparent at visible ...

Detecting and analyzing nanoscale motion patterns of vesicles, smaller than the microscope resolution (~250 nm), inside living biological cells is a challenging problem. State-of-the-art CV approaches based on detection, tracking, optical flow or deep learning perform poorly for this problem. We propose an integrative approach, built upon physics based simulations, nanoscopy algorithms, and shallow ...

The dimensions of mitochondria are close to the diffraction limit of conventional light microscopy techniques, making the complex internal structures of mitochondria unresolvable. In recent years, new fluorescence-based optical imaging techniques have emerged, which allow for optical imaging below the conventional limit, enabling super-resolution (SR). Possibly the most promising SR and diffraction-limited ...

We present an open-source implementation of the fluctuation-based nanoscopy method MUSICAL for ImageJ. This implementation improves the algorithm’s computational efficiency and takes advantage of multi-threading to provide orders of magnitude faster reconstructions than the original MATLAB implementation. In addition, the plugin is capable of generating super-resolution videos from large stacks of ...

Multiple signal classification algorithm (MUSICAL) exploits temporal fluctuations in fluorescence intensity to perform super-resolution microscopy by computing the value of a super-resolving indicator function across a fine sample grid. A key step in the algorithm is the separation of the measurements into signal and noise subspaces, based on a single user-specified parameter called the threshold. ...

Labelfree nanoscopy encompasses optical imaging with resolution in the 100 nm range using visible wavelengths. Here, we present a labelfree nanoscopy method that combines coherent imaging techniques with waveguide microscopy to realize a <i>super-condenser</i> featuring maximally inclined coherent darkfield illumination with artificially stretched wave vectors due to large refractive indices of the ...

Modern science is based on observation, and its advances are limited by what is possible to observe. For centuries optical microscopy was limited by the diffraction of light, making structures closer together than half the wavelength of light unresolvable. In recent years, the new field of optical nanoscopy has emerged, enabling diffraction-limited structures to be resolved. As opposed to ...