Abstract
In this study, we investigate fluorescence optical detection and image reconstruction based on modulated light illumination using a digital micromirror device (DMD). Fluorescence detection is one of the most common methods to study various cellular dynamics labeled with fluorescent indicators. Although employed in many cell-based assays, widefield fluorescence microscopy provides poor axial sectioning capability that is insufficient to measure thick cell-based assays, e.g., those with 3D cell complex cultured in a thick extracellular matrix. Confocal fluorescence microscopy, on the other hand, provides good axial sectioning capability compared to wide-field fluorescence imaging. However, confocal microscopy is subject to temporal overhead associated with scanning to acquire fluorescence images. For image acquisition at enhanced axial sectioning with reduced processing load, we have developed a fluorescence optical detection system based on subtractive light illumination using a DMD. Compared to moving grid masks, a DMD provides fast and flexible scanning by modulating aperture patterns. Here, we report DMD-based structured light illumination for improved image reconstruction by separating in-focus and out-of-focus fluorescence components so that the system can achieve 3D fluorescence image stacks with enhanced axial sectioning capability. In this proof-of-concept study, the DMD-based structured light illumination system was evaluated by observing two-dimensionally deposited fluorescent microbeads and mixed pollen grains. Also shown was that the scanning time to obtain fluorescence images can be greatly reduced compared to confocal microscopy.
| Original language | English |
|---|---|
| Title of host publication | Emerging Digital Micromirror Device Based Systems and Applications V |
| Volume | 8618 |
| DOIs | |
| Publication status | Published - 2013 Jun 12 |
| Event | Emerging Digital Micromirror Device Based Systems and Applications V - San Francisco, CA, United States Duration: 2013 Feb 5 → 2013 Feb 6 |
Other
| Other | Emerging Digital Micromirror Device Based Systems and Applications V |
|---|---|
| Country | United States |
| City | San Francisco, CA |
| Period | 13/2/5 → 13/2/6 |
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All Science Journal Classification (ASJC) codes
- Electronic, Optical and Magnetic Materials
- Condensed Matter Physics
- Computer Science Applications
- Applied Mathematics
- Electrical and Electronic Engineering
Cite this
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Fluorescence image detection and reconstruction by subtractive light illumination using a digital micromirror device. / Choi, Jong Ryul; Kim, Donghyun.
Emerging Digital Micromirror Device Based Systems and Applications V. Vol. 8618 2013. 861808.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
TY - GEN
T1 - Fluorescence image detection and reconstruction by subtractive light illumination using a digital micromirror device
AU - Choi, Jong Ryul
AU - Kim, Donghyun
PY - 2013/6/12
Y1 - 2013/6/12
N2 - In this study, we investigate fluorescence optical detection and image reconstruction based on modulated light illumination using a digital micromirror device (DMD). Fluorescence detection is one of the most common methods to study various cellular dynamics labeled with fluorescent indicators. Although employed in many cell-based assays, widefield fluorescence microscopy provides poor axial sectioning capability that is insufficient to measure thick cell-based assays, e.g., those with 3D cell complex cultured in a thick extracellular matrix. Confocal fluorescence microscopy, on the other hand, provides good axial sectioning capability compared to wide-field fluorescence imaging. However, confocal microscopy is subject to temporal overhead associated with scanning to acquire fluorescence images. For image acquisition at enhanced axial sectioning with reduced processing load, we have developed a fluorescence optical detection system based on subtractive light illumination using a DMD. Compared to moving grid masks, a DMD provides fast and flexible scanning by modulating aperture patterns. Here, we report DMD-based structured light illumination for improved image reconstruction by separating in-focus and out-of-focus fluorescence components so that the system can achieve 3D fluorescence image stacks with enhanced axial sectioning capability. In this proof-of-concept study, the DMD-based structured light illumination system was evaluated by observing two-dimensionally deposited fluorescent microbeads and mixed pollen grains. Also shown was that the scanning time to obtain fluorescence images can be greatly reduced compared to confocal microscopy.
AB - In this study, we investigate fluorescence optical detection and image reconstruction based on modulated light illumination using a digital micromirror device (DMD). Fluorescence detection is one of the most common methods to study various cellular dynamics labeled with fluorescent indicators. Although employed in many cell-based assays, widefield fluorescence microscopy provides poor axial sectioning capability that is insufficient to measure thick cell-based assays, e.g., those with 3D cell complex cultured in a thick extracellular matrix. Confocal fluorescence microscopy, on the other hand, provides good axial sectioning capability compared to wide-field fluorescence imaging. However, confocal microscopy is subject to temporal overhead associated with scanning to acquire fluorescence images. For image acquisition at enhanced axial sectioning with reduced processing load, we have developed a fluorescence optical detection system based on subtractive light illumination using a DMD. Compared to moving grid masks, a DMD provides fast and flexible scanning by modulating aperture patterns. Here, we report DMD-based structured light illumination for improved image reconstruction by separating in-focus and out-of-focus fluorescence components so that the system can achieve 3D fluorescence image stacks with enhanced axial sectioning capability. In this proof-of-concept study, the DMD-based structured light illumination system was evaluated by observing two-dimensionally deposited fluorescent microbeads and mixed pollen grains. Also shown was that the scanning time to obtain fluorescence images can be greatly reduced compared to confocal microscopy.
UR - http://www.scopus.com/inward/record.url?scp=84878727272&partnerID=8YFLogxK
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U2 - 10.1117/12.2003788
DO - 10.1117/12.2003788
M3 - Conference contribution
SN - 9780819493873
VL - 8618
BT - Emerging Digital Micromirror Device Based Systems and Applications V
ER -