ArticleCommunications engineering2025
Paired-objectives photon enhancement (POPE) microscopy: enhanced photon collection for fluorescence imaging.
Article in Communications engineering, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
Fluorescence microscopy is indispensable for visualizing biological structures and dynamics, yet its efficiency is limited-over half of emitted photons fall outside the objective's numerical aperture and go undetected. Here, we introduce Paired-Objectives Photon Enhancement (POPE) microscopy, which increases photon collection efficiency by up to two-fold using a single excitation source, single detector, and dual objectives. By integrating a 4f optical system with a reflective mirror positioned opposite the objective in an inverted microscope, POPE redirects a substantial portion of otherwise lost photons into the detection pathway. Compatible with super-resolution, confocal, epifluorescence, and autofluorescence modalities, POPE improves spatial resolution, acquisition speed, and signal-to-noise ratio, particularly under photon-limited conditions. It has been validated across fluorophore solutions, subcellular structures, live cells, and thick tissues, consistently enhancing imaging performance. As a modular and cost-effective upgrade for standard inverted microscopes, POPE extends access to high-sensitivity fluorescence imaging and enables new applications in cell biology, biophysics, and biomedical research.
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