# Developing multiparametric fluorescence microscopy to connect metabolism with cellular manipulations

> **NIH NIH R35** · GEORGETOWN UNIVERSITY · 2024 · $390,000

## Abstract

Project Summary:
Correlating metabolic changes with external or internal stimuli and cellular manipulations in high spatial detail
is important for understanding causation and correlation in biological systems. Especially, direct pixel-by-pixel
correlation of multiple observable quantities, e.g., metabolism, diffusion of proteins, membrane fluidity, is
difficult to observe simultaneously. Connecting these physical and biophysical behaviors across multiple
imaging modalities can enable us to understand how inhibition of one property, e.g. changing the diffusion of
the protein, can influence other properties such as metabolism or membrane fluidity. This type of correlation
and causation-based simultaneous imaging in the same biological system is something that is lacking in
current fluorescence imaging-based systems. Investigation of these properties in high resolution via confocal
and multiphoton fluorescence microscopes enables us to interrogate cellular and subcellular details of these
interactions. The goal of my research program is to create a framework and seamless workflow of imaging
methodology where all of these fundamental properties and interactions can be quantified and correlated at the
native resolution of the acquired image. To do so, we propose combining the phasor approach to Fluorescence
Lifetime Imaging (phasor-FLIM), phasor approach to hyperspectral imaging (sp-phasor) and phasor approach
to Fluorescence Correlation Spectroscopy (phasor-FCS). We will further create an imaging biophysics
research program which combines single-molecule (SM) and ensemble fluorescence experiments in vitro and
in solution to get a deeper detail at the fundamental questions arising from the molecular interactions. Most
imaging approaches are focused on increasing resolution (either in time or space) for structural elucidation.
Our approach towards imaging is fundamentally different because we are focused on understanding the
functional aspects and quantifying physical properties that can be observed using imaging modalities. In the
next 5 years we want to establish the principle of this multiparametric and multidimensional phasor imaging
and apply them to various biological systems, both in cells and in tissues. Our future goal is to combine the
analysis methods developed here with camera-based systems, including light sheet microscopy, and extend
these approaches to a faster instrumentation and eventually to whole-organism imaging.

## Key facts

- **NIH application ID:** 10938773
- **Project number:** 1R35GM154815-01
- **Recipient organization:** GEORGETOWN UNIVERSITY
- **Principal Investigator:** Suman Ranjit
- **Activity code:** R35 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2024
- **Award amount:** $390,000
- **Award type:** 1
- **Project period:** 2024-09-01 → 2029-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10938773

## Citation

> US National Institutes of Health, RePORTER application 10938773, Developing multiparametric fluorescence microscopy to connect metabolism with cellular manipulations (1R35GM154815-01). Retrieved via AI Analytics 2026-05-25 from https://api.ai-analytics.org/grant/nih/10938773. Licensed CC0.

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