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Updated: Jun 18, 2026
Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice
Published on: January 19, 2018
Multicolor fluorescent intravital live microscopy (FILM) for surgical tumor resection in a mouse xenograft model
Greg M Thurber1, Jose L Figueiredo, Ralph Weissleder
1Center for Systems Biology, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, United States of America.
Background:
Complete surgical resection of neoplasia remains one of the most efficient tumor therapies. However, malignant cell clusters are often left behind during surgery due to the inability to visualize and differentiate them against host tissue. Here we establish the feasibility of multicolor fluorescent intravital live microscopy (FILM) where multiple cellular and/or unique tissue compartments are stained simultaneously and imaged in real time.
Methodology/Principal Findings:
Theoretical simulations of imaging probe localization were carried out for three agents with specificity for cancer cells, stromal host response, or vascular perfusion. This transport analysis gave insight into the probe pharmacokinetics and tissue distribution, facilitating the experimental design and allowing predictions to be made about the localization of the probes in other animal models and in the clinic. The imaging probes were administered systemically at optimal time points based on the simulations, and the multicolor FILM images obtained in vivo were then compared to conventional pathological sections. Our data show the feasibility of real time in vivo pathology at cellular resolution and molecular specificity with excellent agreement between intravital and traditional in vitro immunohistochemistry.
Conclusions/Significance:
Multicolor FILM is an accurate method for identifying malignant tissue and cells in vivo. The imaging probes distributed in a manner similar to predictions based on transport principles, and these models can be used to design future probes and experiments. FILM can provide critical real time feedback and should be a useful tool for more effective and complete cancer resection.
Insights
Multicolor fluorescent intravital live microscopy (FILM) enables real-time, in vivo visualization of cancer cells, improving surgical tumor resection accuracy. This advanced imaging technique aids in differentiating malignant tissue from healthy cells during operations.
Area of Science:
- Biomedical Imaging
- Surgical Oncology
- Molecular Pathology
Background:
- Complete surgical resection is a key cancer therapy, but residual malignant cells often hinder treatment success.
- Current methods struggle to differentiate cancerous cells from healthy tissue during surgery.
- Real-time visualization of neoplastic cells is crucial for improving surgical outcomes.
Purpose of the Study:
- To establish the feasibility of multicolor fluorescent intravital live microscopy (FILM) for in vivo cancer cell detection.
- To develop and validate simultaneous imaging of multiple cellular and tissue compartments.
- To assess the accuracy of FILM in comparison to traditional pathology.
Main Methods:
- Theoretical simulations guided the design of imaging probes targeting cancer cells, stromal response, and vascular perfusion.
- Pharmacokinetic and tissue distribution analyses informed optimal probe administration timing.
- Multicolor FILM was performed in vivo, and images were compared with conventional ex vivo pathological sections.
Main Results:
- Simulations accurately predicted probe localization and tissue distribution.
- In vivo multicolor FILM provided real-time, cellular-resolution imaging with molecular specificity.
- Intravital imaging demonstrated excellent agreement with traditional in vitro immunohistochemistry.
Conclusions:
- Multicolor FILM is a validated method for accurate in vivo identification of malignant tissue and cells.
- Transport modeling aids in designing future imaging probes and experiments.
- FILM offers critical real-time feedback, enhancing the effectiveness of cancer resection surgeries.
