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Dive into the research topics where Birka Hein is active.

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Featured researches published by Birka Hein.


Nature | 2009

Direct observation of the nanoscale dynamics of membrane lipids in a living cell

Christian Eggeling; Christian Ringemann; Rebecca Medda; Günter Schwarzmann; Konrad Sandhoff; Svetlana Polyakova; Vladimir N. Belov; Birka Hein; Claas von Middendorff; Andreas Schönle; Stefan W. Hell

Cholesterol-mediated lipid interactions are thought to have a functional role in many membrane-associated processes such as signalling events. Although several experiments indicate their existence, lipid nanodomains (‘rafts’) remain controversial owing to the lack of suitable detection techniques in living cells. The controversy is reflected in their putative size of 5–200 nm, spanning the range between the extent of a protein complex and the resolution limit of optical microscopy. Here we demonstrate the ability of stimulated emission depletion (STED) far-field fluorescence nanoscopy to detect single diffusing (lipid) molecules in nanosized areas in the plasma membrane of living cells. Tuning of the probed area to spot sizes ∼70-fold below the diffraction barrier reveals that unlike phosphoglycerolipids, sphingolipids and glycosylphosphatidylinositol-anchored proteins are transiently (∼10–20 ms) trapped in cholesterol-mediated molecular complexes dwelling within <20-nm diameter areas. The non-invasive optical recording of molecular time traces and fluctuation data in tunable nanoscale domains is a powerful new approach to study the dynamics of biomolecules in living cells.


Nature Methods | 2008

Fluorescence nanoscopy by ground-state depletion and single-molecule return.

Jonas Fölling; Mariano L. Bossi; Hannes Bock; Rebecca Medda; Christian A. Wurm; Birka Hein; Stefan Jakobs; Christian Eggeling; Stefan W. Hell

We introduce far-field fluorescence nanoscopy with ordinary fluorophores based on switching the majority of them to a metastable dark state, such as the triplet, and calculating the position of those left or those that spontaneously returned to the ground state. Continuous widefield illumination by a single laser and a continuously operating camera yielded dual-color images of rhodamine- and fluorescent protein–labeled (living) samples, proving a simple yet powerful super-resolution approach.


Proceedings of the National Academy of Sciences of the United States of America | 2008

Stimulated emission depletion (STED) nanoscopy of a fluorescent protein-labeled organelle inside a living cell

Birka Hein; Katrin I. Willig; Stefan W. Hell

We demonstrate far-field optical imaging with subdiffraction resolution of the endoplasmic reticulum (ER) in the interior of a living mammalian cell. The diffraction barrier is overcome by applying stimulated emission depletion (STED) on a yellow fluorescent protein tag. Imaging individual structural elements of the ER revealed a focal plane (x, y) resolution of <50 nm inside the living cell, corresponding to a 4-fold improvement over that of a confocal microscope and a 16-fold reduction in the focal-spot cross-sectional area. A similar gain in resolution is realized with both pulsed- and continuous-wave laser illumination. Images of highly convoluted parts of the ER reveal a similar resolution improvement in 3D optical sectioning by a factor of 3 along the optic axis (z). Time-lapse STED recordings document morphological changes of the ER over time. Thus, nanoscale 3D imaging of organelles in the interior of living cells greatly expands the scope of light microscopy in cell biology.


Proceedings of the National Academy of Sciences of the United States of America | 2008

Live-cell imaging of dendritic spines by STED microscopy

U. Valentin Nägerl; Katrin I. Willig; Birka Hein; Stefan W. Hell; Tobias Bonhoeffer

Time lapse fluorescence imaging has become one of the most important approaches in neurobiological research. In particular, both confocal and two-photon microscopy have been used to study activity-dependent changes in synaptic morphology. However, the diffraction-limited resolution of light microscopy is often inadequate, forcing researchers to complement the live cell imaging strategy by EM. Here, we report on the first use of a far-field optical technique with subdiffraction resolution to noninvasively image activity-dependent morphological plasticity of dendritic spines. Specifically we show that time lapse stimulated emission depletion imaging of dendritic spines of YFP-positive hippocampal neurons in organotypic slices outperforms confocal microscopy in revealing important structural details. The technique substantially improves the quantification of morphological parameters, such as the neck width and the curvature of the heads of spines, which are thought to play critical roles for the function and plasticity of synaptic connections.


Nature Methods | 2006

Nanoscale resolution in GFP-based microscopy

Katrin I. Willig; Robert Kellner; Rebecca Medda; Birka Hein; Stefan Jakobs; Stefan W. Hell

We report attainment of subdiffraction resolution using stimulated emission depletion (STED) microscopy with GFP-labeled samples. The ∼70 nm lateral resolution attained in this study is demonstrated by imaging GFP-labeled viruses and the endoplasmic reticulum (ER) of a mammalian cell. Our results mark the advent of nanoscale biological microscopy with genetically encoded markers.


Biochemistry | 2009

A rapidly maturing far-red derivative of DsRed-Express2 for whole-cell labeling.

Rita Strack; Birka Hein; Dibyendu Bhattacharyya; Stefan W. Hell; Robert J. Keenan; Benjamin S. Glick

Fluorescent proteins (FPs) with far-red excitation and emission are desirable for multicolor labeling and live-animal imaging. We describe E2-Crimson, a far-red derivative of the tetrameric FP DsRed-Express2. Unlike other far-red FPs, E2-Crimson is noncytotoxic in bacterial and mammalian cells. E2-Crimson is brighter than other far-red FPs and matures substantially faster than other red and far-red FPs. Approximately 40% of the E2-Crimson fluorescence signal is remarkably photostable. With an excitation maximum at 611 nm, E2-Crimson is the first FP that is efficiently excited with standard far-red lasers. We show that E2-Crimson has unique applications for flow cytometry and stimulated emission depletion (STED) microscopy.


Biophysical Journal | 2010

Stimulated emission depletion nanoscopy of living cells using SNAP-tag fusion proteins.

Birka Hein; Katrin I. Willig; Christian A. Wurm; Volker Westphal; Stefan Jakobs; Stefan W. Hell

We show far-field fluorescence nanoscopy of different structural elements labeled with an organic dye within living mammalian cells. The diffraction barrier limiting far-field light microscopy is outperformed by using stimulated emission depletion. We used the tagging protein hAGT (SNAP-tag), which covalently binds benzylguanine-substituted organic dyes, for labeling. Tetramethylrhodamine was used to image the cytoskeleton (vimentin and microtubule-associated protein 2) as well as structures located at the cell membrane (caveolin and connexin-43) with a resolution down to 40 nm. Comparison with structures labeled with the yellow fluorescent protein Citrine validates this labeling approach. Nanoscopic movies showing the movement of connexin-43 clusters across the cell membrane evidence the capability of this technique to observe structural changes on the nanoscale over time. Pulsed or continuous-wave lasers for excitation and stimulated emission depletion yield images of similar resolution in living cells. Hence fusion proteins that bind modified organic dyes expand widely the application range of far-field fluorescence nanoscopy of living cells.


Optics Express | 2010

Fast STED microscopy with continuous wave fiber lasers.

Gael Moneron; Rebecca Medda; Birka Hein; Arnold Giske; Volker Westphal; Stefan W. Hell

We report on fast beam-scanning stimulated-emission-depletion (STED) microscopy in the visible range using for resolution enhancement compact, low cost and turn-key continuous wave (CW) fiber lasers emitting at 592 nm. Spatial resolutions of 35 to 65 nm in the focal plane are shown for various samples including fluorescent nanoparticles, immuno-stained cells with a non-exhaustive selection of 5 commonly used organic fluorescent markers, and living cells expressing the yellow fluorescent protein Citrine. The potential of the straightforward combination of CW-STED and fast beam scanning is illustrated in a movie of the endoplasmic reticulum (ER) of a living cell, composed of 100 frames (6 microm x 12 microm), each of them acquired in a time shorter than 0.2 s.


Chemistry: A European Journal | 2008

Photostable, amino reactive and water-soluble fluorescent labels based on sulfonated rhodamine with a rigidized xanthene fragment.

Vadim P. Boyarskiy; Vladimir N. Belov; Rebecca Medda; Birka Hein; Mariano L. Bossi; Stefan W. Hell


Biophysical Journal | 2009

High-Resolution Far-Field Fluorescence STED Microscopy Reveals Nanoscale Details of Molecular Membrane Dynamics

Christian Eggeling; Christian Ringemann; Rebecca Medda; Birka Hein; Stefan W. Hell

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