Name: demonstration of a hyperlens-integrated microscope and super-resolution imaging
Uploaded: 2017-09-08T11:45:00
Description: Watch this Scientific Journal Video about Demonstration of a Hyperlens-integrated Microscope and Super-resolution Imaging at JoVE.com

This work is financially supported by Young Investigator program (NRF-2015R1C1A1A02036464), Engineering Research Center program (NRF-2015R1A5A1037668) and Global Frontier program (CAMM-2014M3A6B3063708), M.K., S.S., I.K. acknowledge the Global Ph.D. Fellowships (NRF-2017H1A2A1043204, NRF-2017H1A2A1043322, NRF-2016H1A2A1906519) through the National Research Foundation of Korea (NRF) grant funded by the Ministry of Science, ICT and Future Planning (MSIP) of Korean government.

1. Substrate Preparation
<ol>
	<li>Obtain highly refined quartz wafer. For the fabrication reported here, use a wafer with a 500 &#181;m thickness.</li>
	<li>Spin-coat the quartz wafer with a positive photoresist at 2,000 rpm and bake for 60 s at 90 &#176;C. 
	NOTE: The positive photoresist layer is coated to prevent damage during the subsequent cutting step.</li>
	<li>Use a dicing machine to cut the wafer with photoresist into small pieces 20 x 20 mm2 in size.</li>
	<li>Blow using a compressed nitrogen...

<ol>
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V., Narimanov, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Optical+Hyperlens:+Far-field+imaging+beyond+the+diffraction+limit.">Optical Hyperlens: Far-field imaging beyond the diffraction limit.</a> Opt Express. 14 (18), 8247-8256 (2006).</li><li>Lee, H., Liu, Z., Xiong, Y., Sun, C., Zhang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+optical+hyperlens+for+imaging+below+the+diffraction+limit.">Development of optical hyperlens for imaging below the diffraction limit.</a> Opt Express. 15 (24), 15886-15891 (2007).</li><li>Liu, Z., Lee, H., Xiong, Y., Sun, C., Zhang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Far-Field+Optical+Hyperlens+Magnifying+Sub-Diffraction-Limited+Objects.">Far-Field Optical Hyperlens Magnifying Sub-Diffraction-Limited Objects.</a> Science. 315 (5819), 1686-1686 (2007).</li><li>Kim, M., Rho, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metamaterials+and+imaging.">Metamaterials and imaging.</a> Nano Converg. 2 (1), 22 (2015).</li><li>Byun, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Demonstration+of+nanoimprinted+hyperlens+array+for+high-throughput+sub-diffraction+imaging.">Demonstration of nanoimprinted hyperlens array for high-throughput sub-diffraction imaging.</a> Sci Rep. 7, 46314 (2017).</li><li>Shekhar, P., Atkinson, J., Jacob, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hyperbolic+metamaterials:+fundamentals+and+applications.">Hyperbolic metamaterials: fundamentals and applications.</a> Nano Converg. 1 (1), 14 (2014).</li><li>Liu, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sub-diffraction+demagnification+imaging+lithography+by+hyperlens+with+plasmonic+reflector+layer.">Sub-diffraction demagnification imaging lithography by hyperlens with plasmonic reflector layer.</a> RSC Advances. 6 (98), 95973-95978 (2016).</li><li>Sun, J., Xu, T., Litchinitser, N. 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The fabrication of a hyperlens includes three major steps: defining hemispherical geometry into the quartz substrate through a wet-etching process, stacking the metal and dielectric multilayer using an electron beam evaporation system, and inscribing the object on the Cr layer. The most important step is the second, since it can significantly affect quality of the hyperlens. In the thin-film deposition process, there are two conditions that require special care for a clear super-resolved image. Stacking the multilayer co...

A desire to observe biomolecules in living cells led to the invention of microscopy, and the advent of microscopy propagated the revolution of various fields, such as biology, pathology, and material science, over last few centuries. However, further advancement of research has been restricted by diffraction, which limits the resolution of conventional microscopes to about half of the wavelength1. Therefore, super-resolution imaging to overcome the diffraction limit has been an interesting research area in recent decades.
As the diffraction limit is attributed to the loss of the evanescent waves that contain sub-wavelength information on objects, early studies have been conducted to keep evanescent waves from fading away or to recover them2,3. The effort to overcome the diffraction limit was first reported with near-field scanning optical microscopy, which collects the evanescent field in close proximity to the object before it is dissipated2. However, as scanning the whole image region and reconstructing it takes a long time, it cannot be applied to real-time imaging. Although another approach based on the &#34;superlens,&#34; which amplifies evanescent waves, provides the possibility of real-time imaging, sub-wavelength imaging is only capable in the near-field region and cannot reach far beyond the objects4,5,6,7.
Recently, the hyperlens has emerged as a novel approach to real-time far-field optical imaging8,9,10,11,12. The hyperlens, which is made of highly anisotropic hyperbolic metamaterials13, exhibits a flat hyperbolic dispersion so that it supports high spatial information with the same phase velocity. Furthermore, due to the momentum conservation law, the high transverse wavevector is gradually compressed as the wave goes through the cylindrical geometry. This magnified information thus can be detected by a conventional microscope in the far-field region. This is of particular importance to real-time far-field imaging, as it does not require any point-by-point scanning or image reconstruction. Moreover, the hyperlens can be used for applications other than imaging, including nanolithography. Light that passes through the hyperlens in the reverse direction will be focused onto a sub-diffraction area due to the time-reversal symmetry14,15,16.
Here, we report on a spherical hyperlens that magnifies two-dimensional information at the visible frequency. Unlike conventional cylindrical geometry, the spherical hyperlens magnifies objects in two lateral dimensions, facilitating practical imaging applications. The fabrication method and imaging setup with the hyperlens are presented in detail for the reproduction of a high-quality hyperlens. A sub-wavelength object is inscribed on the hyperlens for the sake of proving its super-resolving power. It is confirmed that small features of inscribed objects are magnified by the hyperlens. Thus, clearly resolved images are obtained in the far-field region in real time. This new type of spherical hyperlens, with its ease of integration with conventional microscopy, provides the possibility of practical imaging applications, leading to the dawn of a new era in biology, pathology, and general nanoscience.

<table>
	<tbody>
		<tr>
			<td>Focused Ion Beam milling machine</td>
			<td>FEI</td>
			<td>Helios Nanolab G3 CX</td>
			<td></td>
		</tr>
		<tr>
			<td>E-beam evaporation system</td>
			<td>Korea Vacuum Tech</td>
			<td>KVE-E4000</td>
			<td></td>
		</tr>
		<tr>
			<td>Scanning electron microscopy</td>
			<td>Hitachi</td>
			<td>SU6600</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted microscopy</td>
			<td>Zeiss</td>
			<td>Axiovert 200</td>
			<td></td>
		</tr>
		<tr>
			<td>Light source</td>
			<td>EXCELITAS Technologies</td>
			<td>X-Cite 110 LED</td>
			<td></td>
		</tr>
		<tr>
			<td>Band pass filter</td>
			<td>Chroma</td>
			<td>ET405/30M</td>
			<td></td>
		</tr>
		<tr>
			<td>Objective lens</td>
			<td>Zeiss</td>
			<td>Plan-Apochromat</td>
			<td>NA=1.3, 100X</td>
		</tr>
		<tr>
			<td>CCD camera</td>
			<td>Andor</td>
			<td>Zyla 4.2</td>
			<td></td>
		</tr>
		<tr>
			<td>Quartz wafer</td>
			<td>CORNING</td>
			<td>Fused Silica Corning 7980</td>
			<td></td>
		</tr>
		<tr>
			<td>Buffered oxide etchant</td>
			<td>J.T Baker TM</td>
			<td>J.T.Baker 5175</td>
			<td></td>
		</tr>
		<tr>
			<td>Photoresist</td>
			<td>AZ electronic materials</td>
			<td>GXR-601 PR</td>
			<td></td>
		</tr>
		<tr>
			<td>Chromium etchant</td>
			<td>SIGMA-ALDRICH</td>
			<td>651826</td>
			<td></td>
		</tr>
		<tr>
			<td>Aceton</td>
			<td>J.T Baker TM</td>
			<td>UN1090</td>
			<td></td>
		</tr>
		<tr>
			<td>Isopropyl alcohol</td>
			<td>J.T Baker TM</td>
			<td>UN1219</td>
			<td></td>
		</tr>
		<tr>
			<td>FEM simulation tool</td>
			<td>COMSOL 5.1 Multiphysics</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

demonstration of a hyperlens-integrated microscope and super-resolution imaging

The use of super-resolution imaging to overcome the diffraction limit of conventional microscopy has attracted the interest of researchers in biology and nanotechnology. Although near-field scanning microscopy and superlenses have improved the resolution in the near-field region, far-field imaging in real-time remains a significant challenge. Recently, the hyperlens, which magnifies and converts evanescent waves into propagating waves, has emerged as a novel approach to far-field imaging. Here, we report the fabrication of a spherical hyperlens composed of alternating silver (Ag) and titanium oxide (TiO2) thin layers. Unlike a conventional cylindrical hyperlens, the spherical hyperlens allows for two-dimensional magnification. Thus, incorporation into conventional microscopy is straightforward. A new optical system integrated with the hyperlens is proposed, allowing for a sub-wavelength image to be obtained in the far-field region in real time. In this study, the fabrication and imaging setup methods are explained in detail. This work also describes the accessibility and possibility of the hyperlens, as well as practical applications of real-time imaging in living cells, which can lead to a revolution in biology and nanotechnology.

The ability of the hyperlens device to resolve sub-diffraction features relies on its uniformity and on a high-quality fabrication. Here, a hyperlens is composed of a multilayer of Ag and TiO2 deposited alternately. Figure 2a shows the SEM image of a well-made hyperlens17. The cross-sectional image shows that the multilayer of Ag and Ti3O5 thin film is deposited with uniform thickness on the hemispheric...

Watch this Scientific Journal Video about Demonstration of a Hyperlens-integrated Microscope and Super-resolution Imaging at JoVE.com

Demonstration of a Hyperlens-integrated Microscope and Super-resolution Imaging

The use of a hyperlens has been regarded as a novel super-resolution imaging technique due to its advantages in real-time imaging and its simple implementation with conventional optics. Here, we present a protocol describing the fabrication and imaging applications of a spherical hyperlens.

The use of super-resolution imaging to overcome the diffraction limit of conventional microscopy has attracted the interest of researchers in biology and ...

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