Two-color 3D MINFLUX movie revealing an inner and outer mitochondrial membrane marker. Cultured mammalian cells labeled with indirect immunofluorescence using JIR AffiniPure-VHH Fragment antibodies (secondary nanobodies) coupled to abberior FLUX 640 (orange) and FLUX 680 (cyan). MINFLUX enables the visualization and separation of both structures.

2D MINFLUX nanoscopy of the nuclear pore complex subunits, labeled with abberior FLUX 647 conjugated to JIR AffiniPure-VHH Fragment antibodies (secondary nanobodies). In contrast to confocal microscopy, 2D MINFLUX allows visualization of the shape and arrangement of individual nuclear pore complex subunits. Here, we reach localization precisions of _~ 2 nm in raw localization data.

Two-color 3D MINFLUX revealing an inner and outer mitochondrial membrane marker. Cultured mammalian cells labeled with indirect immunofluorescence using JIR AffiniPure-VHH Fragment antibodies (secondary nanobodies) coupled to abberior FLUX 640 (orange) and FLUX 680 (cyan). MINFLUX enables the visualization and separation of both structures.

Two color live-cell confocal and STED image of a mammalian cell directly labeled with abberior LIVE 510 mito (cyan), and LIVE RED tubulin. This image was acquired with a STEDYCON microscope.

A brain section stained for the neuronal proteins spectrin and adducine.

Overview image of the zebrafish olfactory epithelium.

Paraffin section of gut biopsy stained for Ki67 (abberior STAR ORANGE), Muc2 (abberior STAR RED), and DAPI.

Gain in both signal-to-background ratio and resolution: MATRIX detection dramatically improves a conventional STED image of the zebrafish olfactory epithelium, resulting in a perfect and crystal-clear image revealing even more detail than STED alone can.

xz section of a stage 17 Drosophila embryo stained for chitin (abberior LIVE 610, green) and DNA (abberior LIVE 550, cyan).

RAYSHAPE preserves resolution and brightness over the whole sample depth of about 200 µm by dynamically redirecting aberrated light to the right places.

In comparison, mechanical optics using a correction collar can only correct a limited z-range of approximately 20 µm

Drosophila stage 12 embryo, imaged with RAYSHAPE, stained for tubulin with abberior STAR RED and for DNA with abberior LIVE 550.

Deep tissue imaging with RAYSHAPE of a stage 17 Drosophila embryo stained for chitin with abberior LIVE 610.

Clear and detailed 3D rendering of trachea imaged with RAYSHAPE and MATRIX STED. Chitin was stained with abberior LIVE 610.

3D STED xz section of a Drosophila embryo trachea, imaged with RAYSHAPE and without abberation correction, depth 15 µm. Chitin was stained with abberior LIVE 610.

2D MINFLUX nanoscopy of Y. enterocolitica sorting platform protein Halo-YscL.

Sample from Alexander Carsten, Prof. Martin Aepfelbacher, Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg Eppendorf, Germany

STED-PAINT imaging of bacterial membranes (magenta) and DNA (green) using a high concentration of the exchangeable labels.

3D MINFLUX nanoscopy of Y. enterocolitica sorting platform protein Halo-YscL in xy view of two bacteria. The z-axis is color coded.

Sample from Alexander Carsten, Prof. Martin Aepfelbacher, Institute of Medical Microbiology, Virology and Hygiene, University Medical Center Hamburg Eppendorf, Germany

Movement of fluorophore-labeled 30S ribosomes (colored) in two E. coli bacteria (grayscale) tracked with MINFLUX.

abberior STAR RED was coupled to polyclonal secondary nanobodies (alpaca VHH single domain antibodies) from Jackson ImmunoResearch and used to label tubulin in fixed mammalian cells via indirect immunofluorescence. The STED image was acquired with the STEDYCON system.