In vivo detection of cryosurgery using multiphoton and harmonic generation microscopy
Related Experiment Video
Updated: Jul 6, 2026
Simultaneous Label-Free Autofluorescence Multi-Harmonic Microscopy
Published on: August 29, 2025
Te-Hsuen Chen1, Chi-Jer Charles Huang, Yi-You Huang
1Institute of Biomedical Engineering, College of Engineering, College of Medicine, National Taiwan University, Taipei 100, Taiwan, ROC.
Abstract:
The tissue injury and the organization of collagen during cryosurgery are poorly characterized because of the lack of appropriate methodologies. In this study, we use multimodal multiphoton microscopy to assess the change of extracellular matrix after cryotreatment of skin. The cellular matrix transformations and the intercellular interactions during the wound healing process after cryolesion for mice were measured in vivo and in real-time through the dorsal skinfold chamber (DSC). Intrinsic second-harmonic generation (SHG) signals from fibrillar collagen and two-photon excited (TPE) autofluorescence from cell were collected to investigate the cryosurgical response in vivo. The TPE and SHG signals are significantly different between normal and cryotreated mice, and correlates with the wound healing process. The results suggest that this approach may be applied in real-time to noninvasively monitor the cryosurgery process and could potentially be applied to clinical evaluation.
Insights
Multimodal multiphoton microscopy noninvasively monitors skin extracellular matrix changes after cryosurgery. This technique tracks collagen and cell alterations, aiding wound healing assessment and potential clinical cryosurgery monitoring.
Area of Science:
- Biomedical Optics
- Dermatology
- Tissue Engineering
Background:
- Cryosurgery's effects on skin tissue and collagen organization lack detailed characterization due to limited methodologies.
- Understanding extracellular matrix dynamics post-cryosurgery is crucial for effective wound healing assessment.
Purpose of the Study:
- To employ multimodal multiphoton microscopy for real-time, in vivo assessment of extracellular matrix changes following cryotreatment of mouse skin.
- To investigate cellular matrix transformations and intercellular interactions during wound healing post-cryolesion.
Main Methods:
- Utilized multimodal multiphoton microscopy in a dorsal skinfold chamber (DSC) model for in vivo, real-time measurements.
- Collected intrinsic second-harmonic generation (SHG) signals from fibrillar collagen.
- Acquired two-photon excited (TPE) autofluorescence signals from cells to analyze cryosurgical response.
Main Results:
- Significant differences observed in TPE and SHG signals between normal and cryotreated mouse skin.
- Multiphoton microscopy signals correlated with the progression of the wound healing process.
- Demonstrated the capability to visualize dynamic changes in the extracellular matrix and cellular behavior.
Conclusions:
- Multimodal multiphoton microscopy offers a noninvasive method for real-time monitoring of cryosurgery-induced tissue alterations.
- This approach shows potential for clinical evaluation and monitoring of cryosurgical procedures.
- Provides novel insights into in vivo tissue response and wound healing mechanisms after cryoablation.
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