Resolution Doubling in 3D-STORM Imaging through Improved Buffers

Olivier, Nicolas; Keller, Debora; Gönczy, Pierre; Manley, Suliana

doi:10.1371/journal.pone.0069004

Cited by 182 publications

(177 citation statements)

References 35 publications

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“…The STORM imaging buffer with oxygen-scavenging system was described previously (Olivier et al 2013) and uses millimolar concentrations of polyunsaturated hydrocarbon cyclooctatetraene to boost photon yields during STORM imaging. All reagents were purchased from Sigma-Aldrich.…”

Section: Storm Image Acquisitionmentioning

confidence: 99%

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The telomeric DNA damage response occurs in the absence of chromatin decompaction

et al. 2017

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Telomeres are specialized nucleoprotein structures that protect chromosome ends from DNA damage response (DDR) and DNA rearrangements. The telomeric shelterin protein TRF2 suppresses the DDR, and this function has been attributed to its abilities to trigger t-loop formation or prevent massive decompaction and loss of density of telomeric chromatin. Here, we applied stochastic optical reconstruction microscopy (STORM) to measure the sizes and shapes of functional human telomeres of different lengths and dysfunctional telomeres that elicit a DDR. Telomeres have an ovoid appearance with considerable plasticity in shape. Examination of many telomeres demonstrated that depletion of TRF2, TRF1, or both affected the sizes of only a small subset of telomeres. Costaining of telomeres with DDR markers further revealed that the majority of DDR signaling telomeres retained a normal size. Thus, DDR signaling at telomeres does not require decompaction. We propose that telomeres are monitored by the DDR machinery in the absence of telomere expansion and that the DDR is triggered by changes at the molecular level in structure and protein composition.

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Section: Storm Image Acquisitionmentioning

confidence: 99%

“…The probe used in this experiment was the Cy5-(CCCTAA) 3 PNA probe (Eurogentec, PN-TC055-005), and the DNA was labeled with DAPI. The oxygenscavenging system used for STORM imaging was glucose oxidase/catalase-based and prepared as described previously (Olivier et al 2013). …”

Section: Storm Image Acquisitionmentioning

confidence: 99%

The telomeric DNA damage response occurs in the absence of chromatin decompaction

et al. 2017

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“…E. coli samples were imaged using Scanasyst-Fluid cantilevers dSTORM. For the imaging with Alexa-647, the sample was imaged in the buffer conditions described in Olivier et al 42 2 For the correlative imaging with ATTO-488, the following protocol was used. After AFM imaging, the sample was washed 3× with PBS, leaving 300 μL PBS.…”

Section: Nano Lettersmentioning

confidence: 99%

High-Resolution Correlative Microscopy: Bridging the Gap between Single Molecule Localization Microscopy and Atomic Force Microscopy

Odermatt¹,

Shivanandan²,

Deschout³

et al. 2015

Nano Lett.

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Nanoscale characterization of living samples has become essential for modern biology. Atomic force microscopy (AFM) creates topological images of fragile biological structures from biomolecules to living cells in aqueous environments. However, correlating nanoscale structure to biological function of specific proteins can be challenging. To this end we have built and characterized a correlated single molecule localization microscope (SMLM)/AFM that allows localizing specific, labeled proteins within high-resolution AFM images in a biologically relevant context. Using direct stochastic optical reconstruction microscopy (dSTORM)/AFM, we directly correlate and quantify the density of localizations with the 3D topography using both imaging modalities along (F-)actin cytoskeletal filaments. In addition, using photo activated light microscopy (PALM)/AFM, we provide correlative images of bacterial cells in aqueous conditions. Moreover, we report the first correlated AFM/PALM imaging of live mammalian cells. The complementary information provided by the two techniques opens a new dimension for structural and functional nanoscale biology.

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“…In order to acquire such images, we prepared biological samples (fixed mammalian cells or purified basal bodies, see Materials and Methods) that were then immersed in a STORM buffer complemented with 2mM Cyclooctatetraene (COT) to enhance the blinking properties of the Alexa 647 fluorophore [8]. Given the high irradiance needed for STORM imaging, samples were first imaged using the SIM mode.…”

Section: Image Acquisition and Reconstructionmentioning

confidence: 99%

Correlative multicolor 3D SIM and STORM microscopy

Hamel

Guichard

Fournier

et al. 2014

Biomed. Opt. Express

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Within the last decade, super-resolution methods that surpass the diffraction limit of light microscopy have provided invaluable insight into a variety of biological questions. Each of these approaches has inherent advantages and limitations, such that their combination is a powerful means to transform them into versatile tools for the life sciences. Here, we report the development of a combined SIM and STORM setup that maintains the optimal resolution of both methods and which is coupled to image registration to localize biological structures in 3D using multicolor labeling. We utilized this workflow to determine the localization of Bld12p/CrSAS-6 in purified basal bodies of Chlamydomonas reinhardtii with utmost precision, demonstrating its usefulness for accurate molecular mapping in 3D. Kihara, Y. Nishida, S. Moriya, M. O. Steinmetz, Y. Hongoh, and P. Gönczy, "Native architecture of the centriole proximal region reveals features underlying its 9-fold radial symmetry," Curr.

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Resolution Doubling in 3D-STORM Imaging through Improved Buffers

Cited by 182 publications

References 35 publications

The telomeric DNA damage response occurs in the absence of chromatin decompaction

The telomeric DNA damage response occurs in the absence of chromatin decompaction

High-Resolution Correlative Microscopy: Bridging the Gap between Single Molecule Localization Microscopy and Atomic Force Microscopy

Correlative multicolor 3D SIM and STORM microscopy

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