Multiscale fluorescent tracking of immune cells in the liver with a highly biocompatible far-red emitting polymer probe - INRAE - Institut national de recherche pour l’agriculture, l’alimentation et l’environnement Accéder directement au contenu
Article Dans Une Revue Scientific Reports Année : 2020

Multiscale fluorescent tracking of immune cells in the liver with a highly biocompatible far-red emitting polymer probe

Résumé

The development of innovative immune cell therapies relies on efficient cell tracking strategies. For this, multiscale fluorescence-based analyses of transferred cells into the host with complementary techniques, including flow cytometry for high-throughput cell analysis and two-photon microscopy for deep tissue imaging would be highly beneficial. Ideally, cells should be labelled with a single fluorescent probe combining all the properties required for these different techniques. Due to the intrinsic autofluorescence of most tissues and especially the liver, far-red emission is also an important asset. However, the development of far-red emitting probes suitable for two-photon microscopy and compatible with clearing methods to track labelled immune cells in thick samples, remains challenging. A newly-designed water-soluble far-red emitting polymer probe, 19K-6H, with a large Stokes shift, was thus evaluated for the tracking of primary immune CD8 T cells. These cells, prepared from mouse spleen, were efficiently labelled with the 19K-6H probe, which was internalized via endocytosis and was highly biocompatible at concentrations up to 20 μM. Labelled primary CD8 T cells were detectable in culture by both confocal and two-photon microscopy as well as flow cytometry, even after 3 days of active proliferation. Finally, 19K-6H-labelled primary CD8 T cells were injected to mice in a classical model of immune mediated hepatitis. The efficient tracking of the transferred cells in the liver by flow cytometry (on purified non-parenchymal cells) and by two-photon microscopy on 800 μm thick cleared sections, demonstrated the versatility of the 19K-6H probe.
Fichier principal
Vignette du fichier
Article Malo_version finale publiee.pdf (6.07 Mo) Télécharger le fichier
Origine : Publication financée par une institution

Dates et versions

hal-02989735 , version 1 (17-11-2021)

Identifiants

Citer

Malo Daniel, Laurence Dubreil, Romain Fleurisson, Jean-Paul Judor, Timothée Bresson, et al.. Multiscale fluorescent tracking of immune cells in the liver with a highly biocompatible far-red emitting polymer probe. Scientific Reports, 2020, 10 (1), ⟨10.1038/s41598-020-74621-9⟩. ⟨hal-02989735⟩
48 Consultations
27 Téléchargements

Altmetric

Partager

Gmail Facebook X LinkedIn More