Advances in Nonlinear Microscopy for Fast Volumetric Imaging in Biology and Medical Sciences

Nonlinear optical microscopy enables intrinsic optical sectioning, label-free contrast, and deep-tissue imaging, and as such it positions photonics at the core of modern biological discovery and translational medicine. As the life sciences advance toward high-throughput and systems-level measurements across space and time, the demand for fast volumetric imaging continues to grow.

Nonlinear optical interactions, such as multiphoton-excited fluorescence, second-harmonic generation (SHG), and third-harmonic generation (THG), provide molecularly and structurally specific signals while enhancing penetration depth and reducing photodamage. Combined with advances in ultrafast laser sources, beam shaping, adaptive optics, and computational reconstruction, these modalities now enable rapid three-dimensional readouts of structure, function, and metabolism in intact tissues and living organisms. Such capabilities are transforming fields including neuroscience, immunology, developmental biology, and clinical diagnostics, where capturing dynamic processes across volumes is essential.

Recent technological developments in nonlinear optical microscopy, particularly in excitation schemes, scanning strategies, and detection architectures, have significantly improved contrast, imaging depth, and acquisition speed, thereby expanding the applicability of this approach across the life and medical sciences. These advances are driven by novel light sources, advanced detection systems, adaptive optics, and computational imaging methods that collectively push imaging performance well beyond the current state of the art.

This Research Topic, fostered by the Horizon2020 project FAIR CHARM (www.faircharm.eu), highlights innovations in instrumentation design, high-speed 3D imaging, enhanced label-free imaging via harmonic generation and intrinsic optical contrast, as well as biological applications ranging from imaging organoids and in vitro 3D models to tissue engineering and in vivo microscopy. Priority will be given to work that bridges technological innovation with transformative impact in the life sciences, from fundamental cell biology to preclinical research.

We welcome submissions of Original Research, Review, Methods, Mini Review, and Perspective articles. Brief Research Reports and Opinion manuscripts will also be considered when aligned with the scope of this collection and the journal’s Author Guidelines. Authors uncertain about fitting the Research Topic are encouraged to contact the Topic Editorial Team prior to submission.

Article types and fees

This Research Topic accepts the following article types, unless otherwise specified in the Research Topic description:

• Brief Research Report
• Editorial
• FAIR² Data
• General Commentary
• Methods
• Mini Review
• Opinion
• Original Research
• Perspective
• ... View all formats

Articles that are accepted for publication by our external editors following rigorous peer review incur a publishing fee charged to Authors, institutions, or funders.

Article types

This Research Topic accepts the following article types, unless otherwise specified in the Research Topic description:

• Brief Research Report
• Editorial
• FAIR² Data
• General Commentary
• Methods
• Mini Review
• Opinion
• Original Research
• Perspective
• Review
• Technology and Code

Keywords: nonlinear optical microscopy, multiphoton fluorescence, second-harmonic generation, third-harmonic generation, fast volumetric imaging, adaptive optics, ultrafast lasers, computational imaging, label-free contrast, deep-tissue imaging

Important note: All contributions to this Research Topic must be within the scope of the section and journal to which they are submitted, as defined in their mission statements. Frontiers reserves the right to guide an out-of-scope manuscript to a more suitable section or journal at any stage of peer review.