Dendrite with spines of a neuron grown on a coverslip, stained for βII spectrin and imaged in confocal mode and with the MINFLUX module of the MIRAVA POLYSCOPE.

Imaging across scales from diffraction-limited to molecular resolution: ribbon synapses in fixed mouse retina tissue stained for VAMP1 (abberior STAR RED) and CtBP2 (abberior STAR ORANGE) (confocal, MATRIX, STED) or for Bassoon (MINFLUX).

Sample courtesy: Arlene Hirano, PhD, University of California Los Angeles, Los Angeles, CA, USA (confocal, MATRIX, STED), and by Dr. Chad Grabner and Prof. Dr. Tobias Moser, Max Planck Institute for Multidisciplinary Sciences, Göttingen, Germany (MINFLUX).

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

MINFLUX imaging of βII spectrin in a primary hippocampal neuron labeled with abberior FLUX 680 by indirect immunofluorescence. Please note the periodic arrangement of spectrin along the axon.

MINFLUX image of axonal βII spectrin labeled with abberior FLUX 660 in primary hippocampal neurons. Note the periodic arrangement of spectrin along the axon, and the absence of any details in the confocal counterpart image.

MINFLUX 3D imaging of βII spectrin in a primary hippocampal neuron labeled with Alexa Fluor 647 by indirect immunofluorescence. The axial coordinate is color-coded. Please note the periodic arrangement of spectrin along the axon.

3D MINFLUX imaging of βII spectrin in a primary hippocampal neuron labeled with Alexa Fluor 647 by indirect immunofluorescence

Confocal and 2D STED imaging of dendrites in brain slices. GFP-tagged proteins were expressed and immunolabelled using primary antibodies against GFP and secondary antibodies coupled to abberior STAR 635P.

Sample was prepared by O. Kaplan and H. Kawabe @ MPI of Experimental Medicine, Göttingen, Germany.

Primary hippocampal neurons with cytoskeleton proteins labeled (magenta, alpha-Adducin, abberior STAR 635P and green, ßII spectrin, Alexa 594). Imaged with abberior Instruments’ STEDYCON and deconvolved with SVI Huygens optimized for STEDYCON.

Body wall preparation of a 3rd instar Drosophila melanogaster larva. Synapses at the neuromuscular junction are visualized by immunostaining against Bruchpilot (Brp), an integral component of active zones in Drosophila. Images were acquired on a STEDYCON attached to an Olympus IX81. Images were kindly provided by Dr. Nadine Ehmann, Institute of Physiology, Neurophysiology, University of Würzburg.

Immunolabelling of βIV-spectrin, a scaffolding protein of the axon initial segment, using a self-made primary antibody and Alexa 594 secondary antibody. Cryo-section of mouse neocortex cut at 20 µm. Images were acquired using a STEDYCON attached to a Zeiss AxioImager Z2. The sample was kindly provided by Dr. Maren Engelhardt, Institute of Neuroanatomy, Medical Faculty Mannheim, Heidelberg University.

MINFLUX image of axonal bII spectrin in primary hippocampal neurons with < 2 nm resolution. Note the periodic arrangement of spectrin along the axon, and the absence of any details in the confocal counterpart image.

Growth cone at the tip of the axon of a primary hippocampal neuron at 1 day in vitro. Microtubules (Tuj1, abberior STAR 580, red) are bundled in the central-domain suggesting a pausing state. The molecular motor myosin IIB (confocal, Alexa488, blue) is enriched at the transition-zone, along the F-actin arcs. In the peripheral domain actin forms bundles in the filopodia (Phalloidin, abberior STAR 635, green). Sample courtesy: Elisa D'Este, Max Planck Institute for Biophysical Chemistry, Göttingen, Germany.

3-color STED image of primary hippocampal neurons. Please note the characteristic ~190 nm beta II spectrin periodicity along distal axons (green) which is only visible in the STED image. Labelled structures: beta II spectrin (green, abberior STAR 635P), Bassoon (red, abberior STAR 580), Actin cytoskeleton (blue, phalloidin, Oregon Green 488). Imaged with abberior Expert Line with 595nm and 775nm STED laser. Sample was prepared by Elisa D’Este @ MPIBPC, Göttingen.

3-color STED imaging: active zones at the Drosophila larval neuromuscular junction immunostained for Bruchpilot and two other proteins.

Two superresolution channels (magenta, yellow) using a 775nm STED laser & one superresolution channel using a 595nm STED laser.

Samples by M. Lenz & M. Landgraf (University of Cambridge, UK).

Spherical aberrations arising from index mismatch between sample and immersion are corrected for, as well as higher-order, sample-induced aberrations.
Without Adaptive Optics, excitation laser power is typically increased with focus depth in order to compensate loss of signal due to aberrations. Adaptive Optics preserves resolution and brightness deep inside thick samples and enables imaging at low light levels.

Sample courtesy of Amelie Cabirol and Albrecht Haase, University of Trento.

3D superresolution volume image of active zones at the Drosophila larval neuromuscular junction. Immunostaining for Bruchpilot and a second protein.

Recorded using EASY3D at 775nm. Samples by M. Lenz & M. Landgraf (University of Cambridge, UK).