Crisp and clear from head to tail – Zoology

Overview image of the zebrafish olfactory epithelium.

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.

Drosophila ovariole stained with abberior LIVE 560 DNA showing nuclei in different cell types of the egg chamber. Ovaries were dissected from adult female fruit flies and were fixed prior to staining.

Image was acquired with the STEDYCON tiling feature and assembled with the SVI Huygens Stitcher.

Drosophila spermatid tails were stained with abberior LIVE 610 tubulin. Testis were dissected from adult male fruit flies. Live cell imaging experiment was performed on a  INFINITY microscope.

Drosophila male accessory gland stained for F-actin using abberior STAR 580 phalloidin.

Sample was prepared in cooperation with Dr. H. R. Shcherbata at MPl for Biophysical Chemistry, Göttingen, Germany.

Drosophila female reproductive system stained for F-actin (red) with abberior STAR RED phalloidin. abberior STAR ORANGE is highlighting a nuclear pore protein (gray).

Image was acquired with the STEDYCON tiling feature and assembled with the SVI Huygens Stitcher.

Actin stain of mouse inner ear hair cells using abberior STAR RED phalloidin.

Samples were prepared by Dr. Christian Vogl, InnerEarLab, UMG Göttingen, Germany.

Actin stain of mouse inner ear hair cells using abberior STAR RED phalloidin. Zoom in for full effect.

Samples were prepared by Dr. Christian Vogl, InnerEarLab, UMG Göttingen, Germany.

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.

Actin stain of mouse inner ear hair cells using abberior STAR RED phalloidin.

Samples were prepared by Dr. Christian Vogl, InnerEarLab, UMG Göttingen, Germany.

2-color 2D STED image of a cleared adult kidney sample of a rat. Shown is an image of a renal corpuscle showing Nephrin (red, abberior STAR 635P) structures inbetween the Podocin slits (green, AlexaFluor594).

Sample was prepared by D. Unnersjö Jess and H.G. Blom @ KTH Stockholm, Sweden.

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.

Drosophila ovariole stained with abberior LIVE 560 DNA showing nuclei in different cell types of the egg chamber. Ovaries were dissected from adult female fruit flies and were fixed prior to staining.

Image was acquired with the STEDYCON tiling feature and assembled with the SVI Huygens Stitcher.

Drosophila ovariole stained with abberior LIVE 560 DNA showing nuclei in different cell types of the egg chamber. Ovaries were dissected from adult female fruit flies and were fixed prior to staining.

Image was acquired with the STEDYCON tiling feature and assembled with the SVI Huygens Stitcher.

Tiled and stitched image of drosophila testis. Sample by J. Rehman.

Drosophila ovariole stained with abberior STAR RED phalloidin (actin, red) and nuclear pores in STAR ORANGE (cyan) highlighting different cell types of the egg chamber.  Image was acquired with the STEDYCON tiling feature and assembled with the HYGENS stitcher

Drosophila ovariole stained with abberior LIVE 560 DNA, showing the different cell types of the egg chamber. The image was stitched using SVI Huygens.

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).

Mouse inner ear hair cells imaged with superresolution in all three dimensions using EASY3D. Animation was generated directly from raw data. Actin stain (abberior STAR 580 phalloidin, blue) and Whirlin immunostain (abberior STAR 635P secondaries, yellow-red). Scalebar 1 µm.

Samples were prepared by Dr. Christian Vogl, InnerEarLab, UMG Göttingen, Germany.

Adaptive optics EASY3D enables deep imaging of cleared adult kidney samples. Comparison of a XZ-slice deep into a renal corpuscle using conventional 3D STED imaging and an EASY3D imaging. EASY3D allowed imaging up to 80 µm into cleared rat kidney tissue using an oil objective. Without adaptive optics, the mismatch between immersion oil and sample leads to complete signal loss. Labels: Nephrin (red, Abberior STAR635P) and Podocin (green, AlexaFluor594).

Sample was prepared by D. Unnersjö Jess and H.G. Blom @ KTH Stockholm, Sweden.

Movie through a 2-color EASY3D RESCUE STED volume stack of a cleared adult kidney sample of a rat. Shown is a movie of an EASY3D and RESCUE volume stack of a renal corpuscle showing Nephrin (red, abberior STAR 635P) structures inbetween the Podocin slits (green, AlexaFluor594).

Sample was prepared by D. Unnersjö Jess and H.G. Blom @ KTH Stockholm, Sweden.

Surface Rendering of 2-color EASY3D volume of a cleared adult kidney sample of a rat. Shown is a reconstruction of an EASY3D volume stack of a renal corpuscle showing Nephrin (red, abberior STAR 635P) structures inbetween the Podocin slits (green, AlexaFluor594).

Sample was prepared by D. Unnersjö Jess and H.G. Blom @ KTH Stockholm, Sweden.