LED-based multicolor extended resolution transmission fluorescence microscopy.

Abstract

Significance: The multiplexing capabilities of fluorescence imaging are enhanced by its exceptional molecular specificity with diverse fluorescent probes, making it a powerful tool for studying complex biological structures, organization, and functions. Recent advances in super-resolution fluorescence microscopy have further revolutionized our ability to explore biology and related fields. However, current multicolor super-resolution fluorescence imaging systems often come with high costs and bulky designs.

Aim: We present a multicolor extended resolution fluorescence imaging system that uses light-emitting diode to simplify the optical path, make the design more compact, and reduce system costs.

Approach: This multicolor extended resolution fluorescence imaging system is based on structured illumination, utilizing a simple diffraction unit positioned between the light source and the sample in a wide-field microscope. Notably, this design could be easily integrated into standard widefield microscopes as a convenient add-on unit, enabling extended resolution imaging.

Results: Our system demonstrates concurrent extended resolved imaging of three-color microsphere beads and successfully showcases multicolor extended resolution fluorescence imaging of biological tissue samples, revealing intricate structural details.

Conclusions: This system provides a structurally simple, cost-effective alternative to traditional microscopes, offering flexible multicolor extended resolution fluorescence imaging and potential applications in multimodal imaging.