Subwavelength imaging through ion-beam-induced upconversion
Identifieur interne : 000A74 ( Ncbi/Merge ); précédent : 000A73; suivant : 000A75Subwavelength imaging through ion-beam-induced upconversion
Auteurs : Zhaohong Mi [Singapour] ; Yuhai Zhang [Singapour] ; Sudheer Kumar Vanga [Singapour] ; Ce-Belle Chen [Singapour] ; Hong Qi Tan [Singapour] ; Frank Watt [Singapour] ; Xiaogang Liu [Singapour, République populaire de Chine] ; Andrew A. Bettiol [Singapour]Source :
- Nature Communications [ 2041-1723 ] ; 2015.
Abstract
The combination of an optical microscope and a luminescent probe plays a pivotal role in biological imaging because it allows for probing subcellular structures. However, the optical resolutions are largely constrained by Abbe's diffraction limit, and the common dye probes often suffer from photobleaching. Here we present a new method for subwavelength imaging by combining lanthanide-doped upconversion nanocrystals with the ionoluminescence imaging technique. We experimentally observed that the ion beam can be used as a new form of excitation source to induce photon upconversion in lanthanide-doped nanocrystals. This approach enables luminescence imaging and simultaneous mapping of cellular structures with a spatial resolution of sub-30 nm.
Url:
DOI: 10.1038/ncomms9832
PubMed: 26560858
PubMed Central: 4660043
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<front><div type="abstract" xml:lang="en"><p>The combination of an optical microscope and a luminescent probe plays a pivotal role in biological imaging because it allows for probing subcellular structures. However, the optical resolutions are largely constrained by Abbe's diffraction limit, and the common dye probes often suffer from photobleaching. Here we present a new method for subwavelength imaging by combining lanthanide-doped upconversion nanocrystals with the ionoluminescence imaging technique. We experimentally observed that the ion beam can be used as a new form of excitation source to induce photon upconversion in lanthanide-doped nanocrystals. This approach enables luminescence imaging and simultaneous mapping of cellular structures with a spatial resolution of sub-30 nm.</p>
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<xref ref-type="aff" rid="a1">1</xref>
<xref ref-type="author-notes" rid="n1">*</xref>
<contrib-id contrib-id-type="orcid">http://orcid.org/0000-0002-9838-3382</contrib-id>
</contrib>
<contrib contrib-type="author"><name><surname>Zhang</surname>
<given-names>Yuhai</given-names>
</name>
<xref ref-type="aff" rid="a2">2</xref>
<xref ref-type="author-notes" rid="n1">*</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Vanga</surname>
<given-names>Sudheer Kumar</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
<contrib-id contrib-id-type="orcid">http://orcid.org/0000-0002-3192-0526</contrib-id>
</contrib>
<contrib contrib-type="author"><name><surname>Chen</surname>
<given-names>Ce-Belle</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Tan</surname>
<given-names>Hong Qi</given-names>
</name>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Watt</surname>
<given-names>Frank</given-names>
</name>
<xref ref-type="corresp" rid="c1">a</xref>
<xref ref-type="aff" rid="a1">1</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Liu</surname>
<given-names>Xiaogang</given-names>
</name>
<xref ref-type="corresp" rid="c2">b</xref>
<xref ref-type="aff" rid="a2">2</xref>
<xref ref-type="aff" rid="a3">3</xref>
<xref ref-type="aff" rid="a4">4</xref>
<contrib-id contrib-id-type="orcid">http://orcid.org/0000-0003-2517-5790</contrib-id>
</contrib>
<contrib contrib-type="author"><name><surname>Bettiol</surname>
<given-names>Andrew A.</given-names>
</name>
<xref ref-type="corresp" rid="c3">c</xref>
<xref ref-type="aff" rid="a1">1</xref>
<xref ref-type="aff" rid="a5">5</xref>
</contrib>
<aff id="a1"><label>1</label>
<institution>Department of Physics, Centre for Ion Beam Applications, National University of Singapore</institution>
, Singapore 117542,<country>Singapore</country>
</aff>
<aff id="a2"><label>2</label>
<institution>Department of Chemistry, National University of Singapore</institution>
, Singapore 117543,<country>Singapore</country>
</aff>
<aff id="a3"><label>3</label>
<institution>Institute of Materials Research and Engineering, Agency for Science, Technology and Research</institution>
, Singapore 117602,<country>Singapore</country>
</aff>
<aff id="a4"><label>4</label>
<institution>Center for Functional Materials, NUS (Suzhou) Research Institute</institution>
, Suzhou, Jiangsu 215123,<country>China</country>
</aff>
<aff id="a5"><label>5</label>
<institution>Yale-NUS College</institution>
, Singapore 138527,<country>Singapore</country>
</aff>
</contrib-group>
<author-notes><corresp id="c1"><label>a</label>
<email>phywattf@nus.edu.sg</email>
</corresp>
<corresp id="c2"><label>b</label>
<email>chmlx@nus.edu.sg</email>
</corresp>
<corresp id="c3"><label>c</label>
<email>a.bettiol@nus.edu.sg</email>
</corresp>
<fn id="n1"><label>*</label>
<p>These authors contributed equally to this work</p>
</fn>
</author-notes>
<pub-date pub-type="epub"><day>12</day>
<month>11</month>
<year>2015</year>
</pub-date>
<pub-date pub-type="collection"><year>2015</year>
</pub-date>
<volume>6</volume>
<elocation-id>8832</elocation-id>
<history><date date-type="received"><day>27</day>
<month>07</month>
<year>2015</year>
</date>
<date date-type="accepted"><day>07</day>
<month>10</month>
<year>2015</year>
</date>
</history>
<permissions><copyright-statement>Copyright © 2015, Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.</copyright-statement>
<copyright-year>2015</copyright-year>
<copyright-holder>Nature Publishing Group, a division of Macmillan Publishers Limited. All Rights Reserved.</copyright-holder>
<license license-type="open-access" xlink:href="http://creativecommons.org/licenses/by/4.0/"><pmc-comment>author-paid</pmc-comment>
<license-p>This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit <ext-link ext-link-type="uri" xlink:href="http://creativecommons.org/licenses/by/4.0/">http://creativecommons.org/licenses/by/4.0/</ext-link>
</license-p>
</license>
</permissions>
<abstract><p>The combination of an optical microscope and a luminescent probe plays a pivotal role in biological imaging because it allows for probing subcellular structures. However, the optical resolutions are largely constrained by Abbe's diffraction limit, and the common dye probes often suffer from photobleaching. Here we present a new method for subwavelength imaging by combining lanthanide-doped upconversion nanocrystals with the ionoluminescence imaging technique. We experimentally observed that the ion beam can be used as a new form of excitation source to induce photon upconversion in lanthanide-doped nanocrystals. This approach enables luminescence imaging and simultaneous mapping of cellular structures with a spatial resolution of sub-30 nm.</p>
</abstract>
<abstract abstract-type="web-summary"><p><inline-graphic id="i1" xlink:href="ncomms9832-i1.jpg"></inline-graphic>
Combining high-resolution microscopic techniques with luminescent probes is important for biological imaging. Here, Mi <italic>et al</italic>
. demonstrate subwavelength imaging by combining lanthanide-doped upconversion nanocrystals with ionoluminescence, revealing cellular structure and particle spatial distribution at high resolution.</p>
</abstract>
</article-meta>
</front>
<floats-group><fig id="f1"><label>Figure 1</label>
<caption><title>Experimental setup and proposed ionoluminescence mechanism.</title>
<p>(<bold>a</bold>
) Artist's view of the basic experimental setup. The focused beam with a spot size of sub-30 nm features can be achieved using a spaced triplet of compact magnetic quadrupole lenses. A Si surface barrier detector is equipped for measuring the energy loss distribution of the ions. (<bold>b</bold>
) Calculated energy distribution of the ionized electrons by bombarding the MeV α-particles on the lanthanide-doped nanocrystals, showing different cross-sections of the resulting electrons at specific energies. Note that most of the ionized electrons have energies mainly located in the visible and infrared spectral region. (<bold>c</bold>
) Proposed upconversion mechanism under α-beam irradiation. The incident helium ions with energy of <italic>E</italic>
<sub>0</sub>
deposit a certain amount of energy (Δ<italic>E</italic>
) onto the crystal to cause the atomic ionization inside the crystal. Subsequently, the ionized secondary electrons can release their energy, most likely during the electron-hole recombination process and successively transfer the energy to Yb<sup>3+</sup>
and Tm<sup>3+</sup>
. An energy transfer from the excited Yb<sup>3+</sup>
to its neighbouring Tm<sup>3+</sup>
ions then populates the excited states (for example, <sup>3</sup>
H<sub>4</sub>
, <sup>1</sup>
G<sub>4</sub>
and <sup>1</sup>
D<sub>2</sub>
) of Tm<sup>3+</sup>
.</p>
</caption>
<graphic xlink:href="ncomms9832-f1"></graphic>
</fig>
<fig id="f2"><label>Figure 2</label>
<caption><title>Spectroscopic analysis of ionoluminescence.</title>
<p>(<bold>a</bold>
) Scanning electron microscopy (SEM) image of the as-synthesized Yb<sup>3+</sup>
/Tm<sup>3+</sup>
-co-doped NaYF<sub>4</sub>
nanorods under investigation. The inserted high-resolution SEM image shows hexagonal cross-sections of these nanorods. Scale bar, 2 μm. (<bold>b</bold>
) Comparative emission spectra of the NaYF<sub>4</sub>
-based nanorods with different dopant compositions when irradiated with α-particles. (<bold>c</bold>
) The plot of the emission ratio of Tm<sup>3+</sup>
and Yb<sup>3+</sup>
(<italic>I</italic>
<sub>Tm</sub>
/<italic>I</italic>
<sub>Yb</sub>
) as a function of Yb<sup>3+</sup>
doping concentration, supporting the energy transfer between the two lanthanide ions. (<bold>d</bold>
) Optimization of Yb<sup>3+</sup>
doping concentration for maximal emission output in the visible range. The ratio of <italic>I</italic>
<sub>Vis</sub>
/<italic>I</italic>
<sub>total</sub>
represents the percentage of integrated visible emission in the total emission covering the range from 350 to 1,100 nm.</p>
</caption>
<graphic xlink:href="ncomms9832-f2"></graphic>
</fig>
<fig id="f3"><label>Figure 3</label>
<caption><title>Luminescence imaging of NaYF<sub>4</sub>
:Yb/Tm (60/2 mol%) nanorods.</title>
<p>(<bold>a</bold>
) Ionoluminescence image of the as-synthesized nanorods through α-particle excitation. (<bold>b</bold>
) High-magnification ionoluminescence image of a single nanorod as marked in <bold>a</bold>
. (<bold>c</bold>
) The corresponding line-scanning profile extracted from the intensity counting at the region marked in <bold>b</bold>
along the arrow, indicating an imaging resolution of about 28 nm. (<bold>d</bold>
) Photoluminescence image of the same sample taken by using 980 nm laser excitation. (<bold>e</bold>
) High-magnification photoluminescence image of the same nanorod as shown in <bold>b</bold>
. (<bold>f</bold>
) The corresponding line-scanning profile from the image shown in <bold>e</bold>
showing a diffraction-limited resolution of 253 nm associated with conventional upconversion microscopes. (<bold>g</bold>
) Ionoluminescence intensity profile as a function of the accumulated dosage of helium ions showing the considerable iono-bleaching resistance of the nanorods. The inserted images, taken at different time intervals (11, 33 and 66 min), indicate that the emission brightness of the nanorods remains essentially unaltered over time. Scale bars, 500 nm. The error bar represents the standard deviation of luminescence counts obtained from a single nanorod in two separate measurements.</p>
</caption>
<graphic xlink:href="ncomms9832-f3"></graphic>
</fig>
<fig id="f4"><label>Figure 4</label>
<caption><title>Particle-distribution mapping by ionoluminescence imaging.</title>
<p>(<bold>a</bold>
) Transmission electron microscopy image of the as-synthesized NaYF<sub>4</sub>
:Yb/Tm (60/2 mol%) nanoparticles. The inserted histogram shows the size distribution of these nanoparticles. (<bold>b</bold>
) Basic experimental design for structural determination and ionoluminescence imaging of the HeLa cell after uptake of the nanoparticles. The luminescence mapping of the nanoparticles and the 3D rendering of detailed cellular structures can be simultaneously implemented by capturing α-particle-induced photons through a photomultiplier tube (PMT) and by scanning transmission ion microscopy, respectively. (<bold>c</bold>
) Comparative photoluminescence (top left) and ionoluminescence (right) imaging, with the latter clearly showing the ability to resolve single nanoparticles (marked by the arrows as shown in the magnified image). Note that top-left and bottom-left images are taken from the same section of the cell. Scale bars, 1 μm. Note that the photoluminescence image was generated by using a 980-nm-diode laser (<xref ref-type="supplementary-material" rid="S1">Supplementary Fig. 12</xref>
).</p>
</caption>
<graphic xlink:href="ncomms9832-f4"></graphic>
</fig>
</floats-group>
</pmc>
<affiliations><list><country><li>République populaire de Chine</li>
<li>Singapour</li>
</country>
</list>
<tree><country name="Singapour"><noRegion><name sortKey="Mi, Zhaohong" sort="Mi, Zhaohong" uniqKey="Mi Z" first="Zhaohong" last="Mi">Zhaohong Mi</name>
</noRegion>
<name sortKey="Bettiol, Andrew A" sort="Bettiol, Andrew A" uniqKey="Bettiol A" first="Andrew A." last="Bettiol">Andrew A. Bettiol</name>
<name sortKey="Bettiol, Andrew A" sort="Bettiol, Andrew A" uniqKey="Bettiol A" first="Andrew A." last="Bettiol">Andrew A. Bettiol</name>
<name sortKey="Chen, Ce Belle" sort="Chen, Ce Belle" uniqKey="Chen C" first="Ce-Belle" last="Chen">Ce-Belle Chen</name>
<name sortKey="Liu, Xiaogang" sort="Liu, Xiaogang" uniqKey="Liu X" first="Xiaogang" last="Liu">Xiaogang Liu</name>
<name sortKey="Liu, Xiaogang" sort="Liu, Xiaogang" uniqKey="Liu X" first="Xiaogang" last="Liu">Xiaogang Liu</name>
<name sortKey="Tan, Hong Qi" sort="Tan, Hong Qi" uniqKey="Tan H" first="Hong Qi" last="Tan">Hong Qi Tan</name>
<name sortKey="Vanga, Sudheer Kumar" sort="Vanga, Sudheer Kumar" uniqKey="Vanga S" first="Sudheer Kumar" last="Vanga">Sudheer Kumar Vanga</name>
<name sortKey="Watt, Frank" sort="Watt, Frank" uniqKey="Watt F" first="Frank" last="Watt">Frank Watt</name>
<name sortKey="Zhang, Yuhai" sort="Zhang, Yuhai" uniqKey="Zhang Y" first="Yuhai" last="Zhang">Yuhai Zhang</name>
</country>
<country name="République populaire de Chine"><noRegion><name sortKey="Liu, Xiaogang" sort="Liu, Xiaogang" uniqKey="Liu X" first="Xiaogang" last="Liu">Xiaogang Liu</name>
</noRegion>
</country>
</tree>
</affiliations>
</record>
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