Near-infrared needle-based confocal laser endomicroscopy (NIR-nCLE) may identify malignant cells at parenchymal staple lines in real-time

Presented During:

Monday, May 8, 2023: 4:15PM - 4:30PM
Los Angeles Convention Center  
Posted Room Name: 408B  

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Abstract Submission 


Patrick Bou-Samra (1), Austin Chang (1), Gregory Kennedy (1), Feredun Azari (1), Eric Bensen (2), John Santini (2), Sunil Singhal (3)


(1) Perelman School of Medicine at the University of Pennsylvania, Philadelphia, PA, (2) Vergent Bioscience, Minneapolis, MN, (3) Hospital of the University of Pennsylvania, Philadelphia, PA

Submitting Author:

Patrick Bou-Samra    -  Contact Me
Perelman School of Medicine at the University of Pennsylvania


Austin Chang    -  Contact Me
Perelman School of Medicine at the University of Pennsylvania
Gregory Kennedy    -  Contact Me
Perelman School of Medicine at the University of Pennsylvania
Feredun Azari    -  Contact Me
Perelman School of Medicine at the University of Pennsylvania
Eric Bensen    -  Contact Me
Vergent Bioscience
John Santini    -  Contact Me
Vergent Bioscience
*Sunil Singhal    -  Contact Me
Hospital of the University of Pennsylvania

Presenting Author:

Patrick Bou-Samra    -  Contact Me



During wedge resections of ground glass opacities, it can be difficult to ascertain negative margins on the parenchymal staple lines by palpation or visualization alone. Frozen section is prone to human error, time consumptive and often difficult to remove staples real-time. We propose a new technology, near-infrared needle-based confocal laser endomicroscopy (NIR-nCLE) to study the presence of cancer cells along the staple line real time during surgery. Our technology utilizes intraoperative molecular imaging with a targeted fluorescent dye and a NIR confocal probe.

A549 cells (human lung adenocarcinoma cell lines) were co-cultured with a targeted fluorescent probe to cathepsin activity (VGT-309) in an Eppendorf and were compared to a negative and positive control. Fluorescence was measured after 2 hours using Iridium Vision Sense Imaging System (Vision Sense, New York, NY). As a proof of concept, we used normal lung parenchyma from a patient with a negative staple line and injected it with A549-VGT309 cells at two separate locations with 3 x106 and 4.5 x 106 cells. We then translated this data into a murine lung cancer model. Finally, the probe was used over the staple line of a resected tumor from patients who received VGT-309 to assess for any residual disease in real-time. NIR-nCLE probe and ImageJ were used to measure the mean fluorescence intensity (MFI).

Fluoresce intensity in A549-VGT in an Eppendorf was 71.8a.u, compared to 12.84a.u in our negative control, confirming selective fluorescence in cancer cells. In our proof of concept model, the sites injected with A549-VGT had a MFI of 147.55 arbitrary units (a.u) and 145.37a.u respectively, which was significantly higher than the negative staple line mean of 55.4a.u (p<0.001). Finally, in our pilot clinical model (n=3), tumor MFI was 175.42a.u and 218.78a.u compared to 49.2a.u and 54.6a.u respectively in normal lung parenchyma (p<0.001). We also had an MFI of 55.83a.u on normal parenchyma compared to 50.55a.u on a negative staple line margin (p=0.2) (figure 1).


NIR-nCLE can detect VGT-targeted cancer cells along a staple line, providing surgeons with a rapid confirmation of their R0 resection. We hypothesize that this technology can be integrated into the OR workflow to expedite surgical decision making and minimize waiting time on pathology with real-time feedback to the surgeon.


Lung Cancer

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Keywords - General Thoracic

Imaging - Imaging
Lung - Lung Cancer