Significance The significance of ultrafast biophotonics lies in its ability to provide novel tools for exploring complex dynamic processes in living systems.Its extremely short timescales and high-resolution imaging capabilities enhance our understanding of the nature of life and drive advances in biomedicine and interdisciplinary integration.Ultrafast time-resolved technology captures transient changes in biomolecules and cells on femtosecond to picosecond timescales,which are challenging to observe under conventional conditions.Through ultrafast spectroscopy,time-resolved imaging,and related techniques,we can monitor critical events such as photophysical processes,energy transfer,and charge separation in real time,revealing the microscopic mechanisms of life activities.This understanding is crucial for comprehending the fundamental principles of living systems and exploring the molecular basis of disease development.Ultrafast biophotonics holds great promise in biomedicine.For example,in cancer diagnosis and treatment,ultrafast spectroscopy technology accurately distinguishes between the optical characteristics of normal cells and cancer cells,providing a reliable basis for early cancer detection.This technology helps to precisely identify tumor boundaries,improving surgical accuracy and success rates.It also enables real-time monitoring of drug distribution and metabolism in the body,supporting tailored treatment approaches.In drug development,ultrafast biophotonics detects structural changes in biomolecules and intermolecular interactions,which is critical for designing safer and more effective drugs.This technology also accelerates drug screening,improving the efficiency of research and development.In addition,ultrafast biophotonics offers a new perspective for biological research by enabling real-time observation of molecular-level changes during biological processes.This deeper understanding of chemical reactions and metabolic processes within organisms helps unravel mysteries in the life sciences and promises breakthroughs in biology.Research in ultrafast biophotonics has progressed through robust interdisciplinary integration,utilizing insights and methods from physics,chemistry,and biology.This integration requires collaborative efforts across disciplines to achieve significant breakthroughs.By combining principles from these diverse fields,ultrafast biophotonics not only drives innovation within its domain,but also catalyzes advances in related disciplines.Physics contributes fundamental theories and experimental techniques critical to understanding light-matter interactions at ultrafast timescales.Concepts such as nonlinear optics,femtosecond laser spectroscopy,and time-resolved imaging play a key role in elucidating biological processes with unprecedented temporal resolution.The application of these principles enables researchers to study molecular dynamics,protein folding kinetics,and cellular signaling pathways in real time.This capability is essential for unraveling the complexity of biological systems from the molecular to the cellular level.Chemistry provides essential tools for the synthesis of novel biomolecular probes and functional materials tailored for ultrafast biophotonic applications.Advances in synthetic chemistry facilitate the development of fluorescent dyes,quantum dots,and photonic crystals optimized for specific biological imaging modalities.Biology provides invaluable insight into the biological relevance and applications of ultrafast biophotonic techniques.By collaborating with biologists and biomedical scientists,photonics researchers gain access to biological samples,experimental models,and clinical data essential for validating their methods.This collaboration fosters a translational approach in which discoveries made at the bench are rapidly translated into clinical diagnostics and therapeutic strategies.In summary,the interdisciplinary nature of ultrafast biophotonics not only enriches the fundamental understanding across physics,chemistry,and biology,but also drives transformative advances in biomedical research and technology.By harnessing the collective expertise of these disciplines,researchers can address complex biological questions and accelerate innovations that benefit both scientific discovery and clinical practice.As the field continues to evolve,interdisciplinary collaboration will remain critical to shaping its trajectory and opening new frontiers in biophotonics and beyond.Ultrafast biophotonics plays a critical role in the study of living systems,the advancement of biomedical science,and the integration of diverse disciplines.Ongoing technological advances and in-depth research will continue to reveal the immense potential and value of ultrafast biophotonics in various fields.Therefore,summarizing existing research is essential to effectively guide future developments in the field.Progress We review the application and development of ultrafast biophotonics in biological systems.We summarize how ultrafast optical techniques detect processes occurring on picosecond to femtosecond timescales.By combining advanced microscopy imaging techniques with ultrafast methods,high temporal and spatial resolution can be achieved.We begin with the basic principles of pump-probe technology and two-dimensional infrared spectroscopy,highlighting their application to the detection of ultrafast biological processes.We then cover near-field ultrafast pump-probe technology,4D optical microscopy,and 4D electron microscopy,emphasizing their high temporal and spatial resolution capabilities(Fig.3).4D optical microscopy allows long-term,high-resolution observation of mitosis in living cells(Fig.6).Finally,we present various spectroscopic mechanisms resulting from nonlinear optical phenomena induced by ultrafast pulses.These mechanisms underlie multiphoton imaging techniques such as second harmonic generation(SHG)imaging,third harmonic generation(THG)imaging,two-photon fluorescence(2PF)imaging,three-photon fluorescence(3PF)imaging,and coherent anti-Stokes Raman scattering(CARS)spectroscopy.Typically,these imaging techniques can coexist in a multimodal microscope,providing multi-angle observation results for biological tissues(Figs.11 and 12).Conclusions and Prospects As ultrafast techniques are increasingly integrated into biophotonics,the field will expand its research scope from single molecules to cells,tissues,and organs.As an emerging interdisciplinary field at the intersection of physics,chemistry,and biology,ultrafast biophotonics requires ongoing research and collaboration to advance its development and expand its applications in biomedical research and technology.
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