# Correlative nonvital standard histopathology and vital multiphoton prehistopathology

> **NIH NIH R41** · LIVEBX, LLC · 2020 · $155,855

## Abstract

SUMMARY
A common practice in medicine for handling freshly biopsied human tissue is to rapidly put it into a formalin
solution. This “fixation” preserves the tissue from decay, allowing transportation to a (remote) pathology
laboratory for standard histopathology or other biochemical/genetical analyses. However, formalin instantly
“kills” the tissue, so that the biochemical and metabolic functions associated with its vital state are permanently
lost. These functions likely contain diagnostic information not available from standard histopathology, e.g. the
pristine picture of an active tumor microenvironment before the structural and chemical perturbations known in
routine histological sample treatments. Thus, it will be highly beneficial to image “tissue vitality” noninvasively
and rapidly (within minutes) before the fixation, by a novel optical imaging technology. Although numerous
technologies have been developed for this purpose, their diagnostic capabilities independent from standard
histopathology have not been unambiguously demonstrated, due to the lack of a correlative technology to co-
register/co-localize the virtual optical sections of vital tissue (optical imaging) and the actual formalin-fixed
paraffin-embedded sections of treated/nonvital tissue (standard histopathology). As a result, it is often difficult
to justify the additional cost of novel optical imaging technologies in clinical standard-of-care processes.
In this project, we aim to develop correlative microscopy between standard histopathology (operated in a
linear/single-photon optical regime) and a novel optical imaging technology based on multiphoton processes,
i.e. multiphoton pre-histopathology. The latter has been demonstrated in our prior work to visualize multiple
unlabeled endogenous biomolecules and segment a rich set of cellular and extracellular components. The
proposed correlative microscopy will retrace optical sections of fresh ex vivo tissue that were imaged using the
label-free multiphoton pre-histopathology to the corresponding formalin-fixed paraffin-embedded sections with
diverse histological stains. For proof-of-concept demonstration (Phase I), we will use discarded fresh rodent
tissue specimens to mimic freshly biopsied human tissue specimens, and co-register a wide area (1x1 mm2) of
image in the multiphoton pre-histopathology with its counterpart in standard hematoxylin and eosin (H&E)
histology. In a future stage (Phase II), we will expand the co-registration area from 1x1 mm2 to 10x10 mm2 to
accommodate larger sample sizes, extend the stains from H&E to diverse immunohistological stains, and
switch the samples from the meat products to fresh human tissue biopsies in a clinical setting. The successful
outcome of this project will equip pathologists with a real-time, label-free, slide-free, digital, and point-of-care
or point-of-procedure tool to image “tissue vitality”, without affecting the existing histology workflow but with
newly added critical inf...

## Key facts

- **NIH application ID:** 10080978
- **Project number:** 1R41GM139528-01
- **Recipient organization:** LIVEBX, LLC
- **Principal Investigator:** Haohua Tu
- **Activity code:** R41 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2020
- **Award amount:** $155,855
- **Award type:** 1
- **Project period:** 2020-08-10 → 2022-08-09

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10080978

## Citation

> US National Institutes of Health, RePORTER application 10080978, Correlative nonvital standard histopathology and vital multiphoton prehistopathology (1R41GM139528-01). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10080978. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*