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Enhanced light collection in fluorescence microscopy using self-assembled micro-reflectors.

ABSTRACT: In fluorescence microscopy, the signal-to-noise ratio (SNR) of the optical system is directly linked to the numerical aperture (NA) of the microscope objective, which creates detection challenges for low-NA, wide-field and high-throughput imaging systems. Here we demonstrate a method to increase the light collection efficiency from micron-scale fluorescent objects using self-assembled vapor-condensed polyethylene glycol droplets, which act as micro-reflectors for fluorescent light. Around each fluorescent particle, a liquid meniscus is formed that increases the excitation efficiency and redirects part of the laterally-emitted fluorescent light towards the detector due to internal reflections at the liquid-air interface of the meniscus. The three-dimensional shape of this micro-reflector can be tuned as a function of time, vapor temperature, and substrate contact angle, providing us optimized SNR performance for fluorescent detection. Based on these self-assembled micro-reflectors, we experimentally demonstrate ~2.5-3 fold enhancement of the fluorescent signal from 2-10 ?m sized particles. A theoretical explanation of the formation rate and shapes of these micro-reflectors is presented, along with a ray tracing model of their optical performance. This method can be used as a sample preparation technique for consumer electronics-based microscopy and sensing tools, thus increasing the sensitivity of low-NA systems that image fluorescent micro-objects.

SUBMITTER: Gorocs Z

PROVIDER: S-EPMC4470325 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

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Enhanced light collection in fluorescence microscopy using self-assembled micro-reflectors.

Göröcs Zoltán Z McLeod Euan E Ozcan Aydogan A

Scientific reports 20150617

In fluorescence microscopy, the signal-to-noise ratio (SNR) of the optical system is directly linked to the numerical aperture (NA) of the microscope objective, which creates detection challenges for low-NA, wide-field and high-throughput imaging systems. Here we demonstrate a method to increase the light collection efficiency from micron-scale fluorescent objects using self-assembled vapor-condensed polyethylene glycol droplets, which act as micro-reflectors for fluorescent light. Around each f ...[more]

PMID: 26083081

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Project description:This study used available or purchased equipment and an image reconstruction system developed by the college of dentistry to establish a basic self-assembled micro-computed tomography (micro-CT) system. Such a system would be suitable for teaching dental radiology to dental students. Specifically, it could help students to understand the principles governing dental cone-beam computed tomography (CBCT) and provide graduate students with a system for scanning small samples (e.g., individual teeth) during the early stages of research. The self-assembled micro-CT system was constructed using a portable dental X-ray tube, an intraoral digital X-ray detector, a high-precision rotation stage, related bracket accessories, and a notebook computer. Reconstructed images and three-dimensional models of the maxillary right third molar were produced using the self-assembled micro-CT system and an advanced commercially available micro-CT system (Skyscan 2211). Subsequently, the reconstructed images and 3D models produced using the two systems were compared by two senior dentists to determine whether considerable visual differences could be observed. Finally, the signal-to-noise ratio (SNR) was used for quantitative analysis and to compare the systems. Although the self-assembled micro-CT system produced image boundaries that were not as sharp as those of Skyscan 2211, the images were nonetheless remarkably similar. In addition, the two micro-CT systems produced 3D models that were almost identical in appearance and root canal shape. Quantitative analysis revealed that Skyscan 2211 had produced a SNR that was superior to that of the self-assembled micro-CT system, with the difference ranging from 36.77% to 136.22%; enamel, which has a higher density, exhibited lower SNR differences, whereas dentin, which has a lower density, exhibited higher SNR differences. The self-assembled micro-CT system with a resolution of 36 μm was created using a portable dental X-ray tube and an intraoral digital X-ray detector. Although the scanning time was relatively long (~30 min to scan images of a tooth), the images were adequate in the preliminary stage of experiments. More importantly, students were afforded the opportunity to observe the process of assembling and disassembling each component of a micro-CT scanner and thereby achieve a more comprehensive understanding of the principles governing micro-CT and dental CBCT.

Dataset Information

Enhanced light collection in fluorescence microscopy using self-assembled micro-reflectors.

Publications

Enhanced light collection in fluorescence microscopy using self-assembled micro-reflectors.

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