Abstract
A major limitation of spontaneous Raman scattering is its intrinsically weak signals, which makes Raman analysis or imaging of biological specimens slow and impractical for many applications. To address this, we report the development of a novel modulated multifocal detection scheme for simultaneous acquisition of full Raman spectra from a 2-D m × n multifocal array. A spatial light modulator (SLM), or a pair of galvo-mirrors, is used to generate m × n laser foci. Raman signals generated within each focus are projected simultaneously into a spectrometer and detected by a CCD camera. The system can resolve the Raman spectra with no crosstalk along the vertical pixels of the CCD camera, e.g., along the entrance slit of the spectrometer. However, there is significant overlap of the spectra in the horizontal pixel direction, e.g., along the dispersion direction. By modulating the excitation multifocal array (illumination modulation) or the emitted Raman signal array (detection modulation), the superimposed Raman spectra of different multifocal patterns are collected. The individual Raman spectrum from each focus is then retrieved from the superimposed spectra using a postacquisition data processing algorithm. This development leads to a significant improvement in the speed of acquiring Raman spectra. We discuss the application of this detection scheme for parallel analysis of individual cells with multifocus laser tweezers Raman spectroscopy (M-LTRS) and for rapid confocal hyperspectral Raman imaging.
| Original language | English (US) |
|---|---|
| Title of host publication | Progress in Biomedical Optics and Imaging - Proceedings of SPIE |
| Publisher | SPIE |
| Volume | 9328 |
| ISBN (Print) | 9781628414189 |
| DOIs | |
| State | Published - 2015 |
| Event | Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIII - San Francisco, United States Duration: Feb 9 2015 → Feb 11 2015 |
Other
| Other | Imaging, Manipulation, and Analysis of Biomolecules, Cells, and Tissues XIII |
|---|---|
| Country | United States |
| City | San Francisco |
| Period | 2/9/15 → 2/11/15 |
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Keywords
- Confocal
- Hyperspectral Raman imaging
- Laser tweezers Raman spectroscopy
- Modulated multifocal detection
- Multifocal array
- Parallel acquisition
- Raman
ASJC Scopus subject areas
- Atomic and Molecular Physics, and Optics
- Electronic, Optical and Magnetic Materials
- Biomaterials
- Radiology Nuclear Medicine and imaging
Cite this
Parallel acquisition of Raman spectra from a 2D multifocal array using a modulated multifocal detection scheme. / Kong, Lingbo; Chan, James W.
Progress in Biomedical Optics and Imaging - Proceedings of SPIE. Vol. 9328 SPIE, 2015. 93280Z.Research output: Chapter in Book/Report/Conference proceeding › Conference contribution
}
TY - GEN
T1 - Parallel acquisition of Raman spectra from a 2D multifocal array using a modulated multifocal detection scheme
AU - Kong, Lingbo
AU - Chan, James W
PY - 2015
Y1 - 2015
N2 - A major limitation of spontaneous Raman scattering is its intrinsically weak signals, which makes Raman analysis or imaging of biological specimens slow and impractical for many applications. To address this, we report the development of a novel modulated multifocal detection scheme for simultaneous acquisition of full Raman spectra from a 2-D m × n multifocal array. A spatial light modulator (SLM), or a pair of galvo-mirrors, is used to generate m × n laser foci. Raman signals generated within each focus are projected simultaneously into a spectrometer and detected by a CCD camera. The system can resolve the Raman spectra with no crosstalk along the vertical pixels of the CCD camera, e.g., along the entrance slit of the spectrometer. However, there is significant overlap of the spectra in the horizontal pixel direction, e.g., along the dispersion direction. By modulating the excitation multifocal array (illumination modulation) or the emitted Raman signal array (detection modulation), the superimposed Raman spectra of different multifocal patterns are collected. The individual Raman spectrum from each focus is then retrieved from the superimposed spectra using a postacquisition data processing algorithm. This development leads to a significant improvement in the speed of acquiring Raman spectra. We discuss the application of this detection scheme for parallel analysis of individual cells with multifocus laser tweezers Raman spectroscopy (M-LTRS) and for rapid confocal hyperspectral Raman imaging.
AB - A major limitation of spontaneous Raman scattering is its intrinsically weak signals, which makes Raman analysis or imaging of biological specimens slow and impractical for many applications. To address this, we report the development of a novel modulated multifocal detection scheme for simultaneous acquisition of full Raman spectra from a 2-D m × n multifocal array. A spatial light modulator (SLM), or a pair of galvo-mirrors, is used to generate m × n laser foci. Raman signals generated within each focus are projected simultaneously into a spectrometer and detected by a CCD camera. The system can resolve the Raman spectra with no crosstalk along the vertical pixels of the CCD camera, e.g., along the entrance slit of the spectrometer. However, there is significant overlap of the spectra in the horizontal pixel direction, e.g., along the dispersion direction. By modulating the excitation multifocal array (illumination modulation) or the emitted Raman signal array (detection modulation), the superimposed Raman spectra of different multifocal patterns are collected. The individual Raman spectrum from each focus is then retrieved from the superimposed spectra using a postacquisition data processing algorithm. This development leads to a significant improvement in the speed of acquiring Raman spectra. We discuss the application of this detection scheme for parallel analysis of individual cells with multifocus laser tweezers Raman spectroscopy (M-LTRS) and for rapid confocal hyperspectral Raman imaging.
KW - Confocal
KW - Hyperspectral Raman imaging
KW - Laser tweezers Raman spectroscopy
KW - Modulated multifocal detection
KW - Multifocal array
KW - Parallel acquisition
KW - Raman
UR - http://www.scopus.com/inward/record.url?scp=84932082322&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84932082322&partnerID=8YFLogxK
U2 - 10.1117/12.2079661
DO - 10.1117/12.2079661
M3 - Conference contribution
AN - SCOPUS:84932082322
SN - 9781628414189
VL - 9328
BT - Progress in Biomedical Optics and Imaging - Proceedings of SPIE
PB - SPIE
ER -