Non-invasive synchronous monitoring of neutrophil migration using whole body near-infrared fluorescence-based imaging

Abstract

Advances in fluorescence imaging coupled with the generation of near infrared probes have significantly improved the capabilities of non-invasive, real-time imaging in whole animals. In this study we were able to overcome a limitation of in vivo fluorescence imaging and have established a dual cell tracking method where two different cell types can be monitored according to the spectral signature of the cell labelling fluorophore. Using a mouse model of acute liver injury, we have characterised the in vivo migration patterns of wild type and transgenic neutrophils with impaired chemotaxis. Here, we were able to demonstrate that IVIS provides a sensitive multiplexing technology to differentiate two different cell populations based on the spectral signature of the cell labelling fluorophores. This spectral unmixing methodology has the potential to uncover multidimensional cellular interactions involved in many diseases such as fibrosis and cancer. In vivo spectral un-mixing provides a useful tool for monitoring multiple biological process in real-time in the same animal.

Whole-Body Imaging to Assess Cell-Based Immunotherapy: Preclinical Studies with an Update on Clinical Translation

“…However, because luciferin and luciferase are foreign proteins to humans, and therefore immunogenic, BLI is not applicable for humans. Fluorescent imaging with near infra-red probes has been used for tracking ex vivo labeled cells such as T cells and neutrophils [56,57]. Optical imaging is subject to tissue absorption of light, and whole-body quantification is not possible [58].…”

Impact of retrotransposon protein L1 ORF1p expression on oncogenic pathways in hepatocellular carcinoma: the role of cytoplasmic PIN1 upregulation

“…Images were analysed using Living Image (version 4.7.2, Perkin Elmer). In vivo and ex vivo photon signals were quantified using the region of Interest method of analysis [ 38 ]. Harvested organs were fixed in absolute ethanol and then embedded in paraffin for IHC analysis (Supplementary Methods for details).…”

Plasmonic random laser enabled artefact-free wide-field fluorescence bioimaging: uncovering finer cellular features

“…Fluorescence imaging is an indispensable tool in life science research and medical diagnostics owing to its contributions to understanding cell physiology, transport characteristics, and interactions of ions, proteins, and other macromolecules. [1][2][3][4][5][6] Lasers, with their intense, narrow-band, and tunable emission, have paved the way for a variety of novel applications based on uorescence imaging. However, the high coherence of the laser leads to the interference of light scattered from optical surfaces, dust particles, inherent imperfections in the system and the sample surface.…”