Name: lensless fluorescent microscopy on a chip
Uploaded: 2011-08-17T13:00:00
Description: Watch this Scientific Journal Video about Lensless Fluorescent Microscopy on a Chip at JoVE.com

A. Ozcan gratefully acknowledges the support of NSF CAREER Award, the ONR Young Investigator Award 2009 and the NIH Director&#x2019;s New Innovator Award DP2OD006427 from the Office of The Director, NIH. The authors also acknowledge the support of the Bill &amp; Melinda Gates Foundation, Vodafone Americas Foundation, and NSF BISH program (under Awards # 0754880 and 0930501).

In this section, we will review the experimental methods of our lensless on-chip fluorescent microscopy platform1-4. To demonstrate the capabilities of this technique, we will show on-chip imaging results for fluorescent micro-particles and labeled white blood cells. Although not discussed here, the same lensfree fluorescent microscopy platform can also be used to image small model animals such as transgenic C. elegans samples3.
1. Design of the Lensless On-Chip Im...

<ol>
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We demonstrated an on-chip fluorescent microscopy platform that can achieve e.g., &lt;4&mu;m spatial resolution over e.g., &gt;0.6-8 cm2 field-of-view without the use of any lenses, mechanical-scanning or thin-film interference-filters. In this technique, with the use of a fiber-optic faceplate or a taper, the fluorescent emission from the objects is collected with a 2D-array of fiber-optic cables before being delivered to an opto-electronic sensor-array such as a CCD/CMOS chip. These acquired lensfree images ...

<table border="1">
 <tbody>
 <tr>
 <th>Material Name</th>
 <th>Company</th>
 <th>Catalogue number</th>
 </tr>
 <tr>
 <td>Charge-coupled device(CCD) </td>
 <td>KODAK</td>
 <td>KAF-8300</td>
 </tr>
 <tr>
 <td>Charge-coupled device(CCD)</td>
 <td>KODAK</td>
 <td>KAF-11002</td>
 </tr>
 <tr>
 <td>Charge-coupled device(CCD)</td>
 <td>KODAK</td>
 <td>KAF-39000</td>
 </tr>
 <tr>
 <td>Complementary Metal-Oxide-Semiconductor (CMOS)</td>
 <td>Micron</td>
 <td>MT9T031C12STCD</td>
 </tr>
 <tr>
 <td>High power LED light source</td>
 <td>Thorlabs </td>
 <td>M455L2-C2</td>
 </tr>
 <tr>
 <td>High power LED driver</td>
 <td>Thorlabs</td>
 <td>LEDD1B</td>
 </tr>
 <tr>
 <td>Fiber coupled LED light source</td>
 <td>Mightex</td>
 <td>FCS-0625-000</td>
 </tr>
 <tr>
 <td>Vacuum Pen</td>
 <td>Edmund Optics</td>
 <td>NT57-636</td>
 </tr>
 <tr>
 <td>2, 4, 10 &mu;m Fluospheres</td>
 <td>Invitrogen</td>
 <td>F-8826, F-8859, F-8836</td>
 </tr>
 <tr>
 <td>RBS lysis buffer 1X </td>
 <td>eBioscience</td>
 <td>00-4333</td>
 </tr>
 <tr>
 <td>SYTO 16 labeling reagent</td>
 <td>Invitrogen</td>
 <td> S7578</td>
 </tr>
 <tr>
 <td>Fiber-optic faceplate</td>
 <td>Edmund Optics</td>
 <td>NT55-142</td>
 </tr>
 <tr>
 <td>Fiber-optic taper</td>
 <td>Edmund Optics</td>
 <td>NT55-134</td>
 </tr>
 <tr>
 <td>Prisms</td>
 <td>Edmund Optics</td>
 <td>NT47-626, NT45-403</td>
 </tr>
 <tr>
 <td>Filters</td>
 <td>Edmund Optics</td>
 <td>NT39-417</td>
 </tr>
 <tr>
 <td>PDMS Elastomers</td>
 <td>Dow Corning</td>
 <td>Slygard 184</td>
 </tr>
 </tbody>
</table>

lensless fluorescent microscopy on a chip

On-chip lensless imaging in general aims to replace bulky lens-based optical microscopes with simpler and more compact designs, especially for high-throughput screening applications. This emerging technology platform has the potential to eliminate the need for bulky and/or costly optical components through the help of novel theories and digital reconstruction algorithms. Along the same lines, here we demonstrate an on-chip fluorescent microscopy modality that can achieve e.g., &lt;4&mu;m spatial resolution over an ultra-wide field-of-view (FOV) of &gt;0.6-8 cm2 without the use of any lenses, mechanical-scanning or thin-film based interference filters. In this technique, fluorescent excitation is achieved through a prism or hemispherical-glass interface illuminated by an incoherent source. After interacting with the entire object volume, this excitation light is rejected by total-internal-reflection (TIR) process that is occurring at the bottom of the sample micro-fluidic chip. The fluorescent emission from the excited objects is then collected by a fiber-optic faceplate or a taper and is delivered to an optoelectronic sensor array such as a charge-coupled-device (CCD). By using a compressive-sampling based decoding algorithm, the acquired lensfree raw fluorescent images of the sample can be rapidly processed to yield e.g., &lt;4&mu;m resolution over an FOV of &gt;0.6-8 cm2. Moreover, vertically stacked micro-channels that are separated by e.g., 50-100 &mu;m can also be successfully imaged using the same lensfree on-chip microscopy platform, which further increases the overall throughput of this modality. This compact on-chip fluorescent imaging platform, with a rapid compressive decoder behind it, could be rather valuable for high-throughput cytometry, rare-cell research and microarray-analysis.

Watch this Scientific Journal Video about Lensless Fluorescent Microscopy on a Chip at JoVE.com

Lensless Fluorescent Microscopy on a Chip

A lensless on-chip fluorescent microscopy platform is demonstrated that can image fluorescent objects over an ultra-wide field-of-view of e.g., &#62;0.6-8 cm2 with &#60;4&#956;m resolution using a compressive sampling based decoding algorithm. Such a compact and wide-field fluorescent on-chip imaging modality could be valuable for high-throughput cytometry, rare-cell research and microarray-analysis.

On-chip lensless imaging in general aims to replace bulky lens-based optical microscopes with simpler and more compact designs, especially for ...

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