Poster De Conférence Année : 2025

Harmonic imaging for tracking live human stem cells in whole blood at high-flow speed and GHz pixel rate

Ameni Dhouib
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Résumé

Preclinical validation of the safety of cell therapy-based strategies requires precise monitoring of the biodistribution of transplanted cells within the host organism. Indeed, the detection of cells in the circulatory system as well as the identification of any possible aggregation state provide critical insights. However, this task demands deep penetration, fast imaging, high selectivity, subcellular resolution, and high throughput. In this study, we demonstrate and characterize a multiphoton-based in-flow detection method for live human stem cells in whole blood (unfiltered or diluted) using a microfluidic channel. Various experimental conditions are investigated such as laser power, stem cell concentration in blood, and flow rate. The approach is based on nonlinear optical microscopy with diffractive scanning perpendicular to the flow direction, enabled by a rapidly wavelength-swept laser. Stem cells were labeled with lithium niobate (LNO) harmonic nanoparticles (HNPs), which exhibit both second and third harmonic generation (SHG and THG). The combination of this setup and HNP-labeled cells enabled imaging at 4,000 frames per second with a pixel dwell time of 2.5 ns. Automated HNP-labeled cell detection and segmentation allowed extraction of key parameters such as single cell diameter, velocity, and recognition between single cells and aggregates. This combination of high-speed multiphoton microscopy and sensitive HNP labeling of cells in opaque and dense media offers a promising platform for detecting specific cells in the bloodstream under intravital microscopy, paving the way for evaluating the safety and dissemination of novel therapeutic agents.

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Dates et versions

hal-05111674 , version 1 (13-06-2025)

Identifiants

  • HAL Id : hal-05111674 , version 1

Citer

Saavedra-Villanueva Oscar, Leroux Isabelle, Wiggert Moritz, Schuetzeck Lion, Rixius Maximilian, et al.. Harmonic imaging for tracking live human stem cells in whole blood at high-flow speed and GHz pixel rate. Interdisciplinary challenges in the physics of complexity and life 2025 (IntCha25), Jun 2025, Heidelberg, Germany. ⟨hal-05111674⟩
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