# Neutrophil Microscopy and Quantitative Imaging Core B

> **NIH NIH P01** · SCRIPPS RESEARCH INSTITUTE, THE · 2022 · $300,559

## Abstract

SUMMARY
Neutrophils are the first line of defense against bacteria and fungi and induce inflammation. The study of the
molecular mechanisms mediating neutrophil activation is therefore of fundamental importance. Visualizing these
mechanisms requires unique, state-of-the-art technologies and approaches. To achieve this, Core B is designed
to coordinate, design, develop and execute new technical approaches applied to the understanding of neutrophil
biology in inflammation. Core B will work with all three Projects and will provide relevant imaging and microscopy
assays in mice and human samples. The core utilizes available cutting-edge technology to implement new
experimental approaches and to help elucidate the intricate cell biology processes proposed in research plans
of Projects 1 (Dr. Hedrick), Project 2 (Dr. Catz) and Project 3 (Dr. Hoffman). To this end, we will have the
dedicated effort of a Technology Core director (Dr. Kiosses) who will allocate specific time and efforts to support
the goals of the PPG. Core B optimizes the workflows of neutrophil imaging, image quantification and data
sharing for all projects. It provides stringent quality controls and ensures uniformity of performance of multiple
key technologies applied to the study of neutrophils and precursors in the setting of coronary artery disease.
Core B will implement unique live-cell approaches to study neutrophil vesicular trafficking, cytoskeleton
remodeling and cell death pathways. Core B is technically and conceptually innovative because it utilizes,
neutrophil-dedicated, novel approaches that include super-resolution microscopy and quantitative analyses of
neutrophil dynamics to elucidate neutrophil functions and to elucidate their role in inflammation. In relationship
with Project 1, Core B will identify neutrophils and their precursors in atherosclerotic plaques during NETosis
and pyroptosis in conditions of inflammasome activation at various stages of plaque development in high-fat diet,
both ex-vivo and in vivo using the ApoE-/- Western diet model of CAD mice. In association with Project 2, Core
B will utilize single-molecule super-resolution microscopy and Total Internal Reflection Fluorescence Microscopy
to study molecular mechanisms regulating vesicular trafficking and actin remodeling in neutrophil precursors.
Core B will also utilize Correlated-Light Electron Microscopy (CLEM) to characterize the newly identified
precursor-specific granules (PSGs). In collaboration with Project 3, Core B will examine three-dimensional
localization and spatial changes in the mitochondrial network as the cells undergo apoptotic or necroptotic cell
death in both neutrophils and their precursors from mouse models and human neutrophils from patients with
CAD. In conclusion, Core B is uniquely designed to use the valuable technology available in La Jolla to the fullest
extent possible to maximize neutrophil-dedicate research and productivity associated with the PPG.

## Key facts

- **NIH application ID:** 10470239
- **Project number:** 5P01HL152958-02
- **Recipient organization:** SCRIPPS RESEARCH INSTITUTE, THE
- **Principal Investigator:** Bill William Kiosses
- **Activity code:** P01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2022
- **Award amount:** $300,559
- **Award type:** 5
- **Project period:** 2021-08-16 → 2026-05-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10470239

## Citation

> US National Institutes of Health, RePORTER application 10470239, Neutrophil Microscopy and Quantitative Imaging Core B (5P01HL152958-02). Retrieved via AI Analytics 2026-05-22 from https://api.ai-analytics.org/grant/nih/10470239. Licensed CC0.

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