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Updated: Jun 30, 2025
Simultaneous Multicolor Imaging of Biological Structures with Fluorescence Photoactivation Localization Microscopy
Published on: December 9, 2013
Multicolor fluorescence microscopy for surgical guidance using a chip-scale imager with a low-NA fiber optic plate
Micah Roschelle1,2, Rozhan Rabbani1, Efthymios Papageorgiou1
1Department of Electrical Engineering and Computer Sciences, University of California, Berkeley, California 94720, USA.
Abstract:
In curative-intent cancer surgery, intraoperative fluorescence imaging of both diseased and healthy tissue can help to ensure the successful removal of all gross and microscopic diseases with minimal damage to neighboring critical structures, such as nerves. Current fluorescence-guided surgery (FGS) systems, however, rely on bulky and rigid optics that incur performance-limiting trade-offs between sensitivity and maneuverability. Moreover, many FGS systems are incapable of multiplexed imaging. As a result, clinical FGS is currently limited to millimeter-scale detection of a single fluorescent target. Here, we present a scalable, lens-less fluorescence imaging chip, VISION, capable of sensitive and multiplexed detection within a compact form factor. Central to VISION is a novel optical frontend design combining a low-numerical-aperture fiber optic plate (LNA-FOP) and a multi-bandpass interference filter, which is affixed to a custom CMOS image sensor. The LNA-FOP acts as a planar collimator to improve resolution and compensate for the angle-sensitivity of the interference filter, enabling high-resolution and multiplexed fluorescence imaging without lenses. We show VISION is capable of detecting tumor foci of less than 100 cells at near video framerates and, as proof of principle, can simultaneously visualize both tumors and nerves in ex vivo prostate tissue.
Insights
A new lens-less fluorescence imaging chip, VISION, enables sensitive, multiplexed detection for cancer surgery. This compact system visualizes both tumors and nerves, improving disease removal and preserving critical structures.
Area of Science:
- Medical Imaging
- Optical Engineering
- Cancer Surgery
Background:
- Intraoperative fluorescence imaging aids cancer surgery by distinguishing diseased from healthy tissue.
- Current fluorescence-guided surgery (FGS) systems have limitations in sensitivity, maneuverability, and multiplexed imaging.
- Existing FGS technology is restricted to detecting single targets at millimeter scales.
Purpose of the Study:
- To develop a scalable, lens-less fluorescence imaging chip for improved intraoperative guidance.
- To overcome the trade-offs between sensitivity and maneuverability in current FGS systems.
- To enable sensitive, multiplexed fluorescence detection in a compact form factor.
Main Methods:
- Designed a novel optical frontend combining a low-numerical-aperture fiber optic plate (LNA-FOP) and a multi-bandpass interference filter.
- Integrated the optical frontend with a custom CMOS image sensor for a lens-less design.
- Utilized the LNA-FOP as a planar collimator to enhance resolution and filter angle-sensitivity.
Main Results:
- The VISION chip achieves sensitive and multiplexed fluorescence imaging without lenses.
- Demonstrated detection of tumor foci comprising fewer than 100 cells at near video framerates.
- Successfully visualized both tumors and nerves simultaneously in ex vivo prostate tissue.
Conclusions:
- The lens-less VISION chip offers a compact and high-performance solution for fluorescence-guided surgery.
- This technology has the potential to improve the accuracy of tumor resection and minimize damage to surrounding tissues.
- VISION enables simultaneous visualization of multiple fluorescent targets, advancing the capabilities of intraoperative imaging.
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01:16Confocal Fluorescence Microscopy