# Probe-based two photon microscopy for functional, label-free early cancer diagnosis

> **NIH NIH R01** · TUFTS UNIVERSITY MEDFORD · 2020 · $747,020

## Abstract

Improvements in the detection of cancerous changes at the pre-invasive stage have the potential to make a
significant impact in the prognosis and treatment of most cancer patients. Despite important advances in our
understanding of cancer pathobiology, the most prevalent diagnostic methods continue to rely on low
magnification tissue visualization, followed by biopsy and histopathology. This process is invasive, often limited
in its sensitivity and/or specificity, time-consuming, and relies heavily on the expertise of highly trained
physicians, who in turn exploit primarily morphological tissue changes for their assessments. These limitations
present barriers to effective treatment and raise monetary and psychological costs. Our long-term objective is to
transform pre- and early epithelial cancer diagnosis through the use of functional (metabolic) and morphological
tissue biomarkers that are extracted non-invasively, automatically, and in near real time from fiber-probe-based
endogenous two-photon (2P) images. In this proposal, we aim to establish and validate such measurements and
biomarkers for the detection of human cervical pre-cancers in vivo. The cervix is an ideal organ for developing
and testing our approach as it relaxes some of the size limitations presented for endoscopic applications,
enabling us to focus on demonstrating the principles of this innovative platform. In addition, we expect that our
proposed methods will enable significant near-term improvements in the sensitivity and specificity of detection
of cervical pre-cancerous lesions during colposcopy and triage with therapy. To achieve our goals, we have
forged a strong partnership among colleagues in academia and the clinic, leveraging strengths and expertise of
multiple teams. Specifically, we will exploit our experience in laser development (Xu, Cornell) and multiphoton
imaging probe design (Ben-Yakar, UT Austin) to develop a probe-based 2P imaging system that is portable and
enables fast image acquisition throughout the cervical epithelium depth with submicron resolution (Aim 1). The
final design specifications will be optimized to enable automated, near-real time analysis of images that provides
quantitative metrics of metabolic function and morphology (Aim 2). In particular, we will assess: a) multiple
quantitative biomarkers of cellular metabolism based on redox ratio and mitochondrial organization parameters
extracted from endogenous NAD(P)H and FAD 2P excited fluorescence images of the epithelium, and b)
morphological metrics associated with depth-dependent variations of the nuclear to cytoplasmic area ratio and
epithelial thickness (Georgakoudi, Tufts). The real time algorithms will be established using freshly excised
normal and pre-cancerous human cervical tissues (Aim 2). They will be tested and further optimized in vivo when
the innovative 2P imaging platform will be used to perform the first-in-human 2P probe-based imaging during
colposcopy (Thieu/Genega, Tufts M...

## Key facts

- **NIH application ID:** 10030979
- **Project number:** 1R01EB030061-01
- **Recipient organization:** TUFTS UNIVERSITY MEDFORD
- **Principal Investigator:** ADELA BEN-YAKAR
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $747,020
- **Award type:** 1
- **Project period:** 2020-07-01 → 2024-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10030979

## Citation

> US National Institutes of Health, RePORTER application 10030979, Probe-based two photon microscopy for functional, label-free early cancer diagnosis (1R01EB030061-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10030979. Licensed CC0.

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