Label-Free In-Line Characterization of Immune Cell Culture using Quantitative Phase Imaging
This article has been Reviewed by the following groups
Discuss this preprint
Start a discussion What are Sciety discussions?Listed in
- Evaluated articles (Arcadia Science)
Abstract
Cell therapies, including T cell immunotherapies, offer promising treatments for previously untreatable diseases, but their widespread use is hindered by challenges in monitoring therapeutic cells during culture -- impacting consistency, potency, and cost. This work demonstrates the use of quantitative phase imaging (QPI), specifically a compact, non-interferometric form called quantitative oblique back illumination microscopy (qOBM), for non-destructive, label-free, in-line assessment of T cell cultures. qOBM enables near real-time feedback on culture growth, contamination, and cell status (viability and activation), comparable to flow cytometry. We further apply this method to characterize genetically modified CAR T cells and explore its potential for advanced T cell phenotyping. Analysis of data from over 50 independent donors shows strong correlation between qOBM metrics and traditional destructive at-line assays. Overall, qOBM provides a powerful tool for continuous, in-line monitoring of therapeutic cell cultures, which can be transformative for improving reproducibility, reducing costs, and advancing the development of cell-based therapies.
Article activity feed
-
From these images, we obtain a pixel-wise dynamic frequency response given by the absolute value of the Fourier transform of the temporal phase signal,
for each spatial pixel in the image.Just to clarify: are these the absolute pixels in the entire imaging field? or the pixels of individual segmented cells? if segmented cells, was there any registration of the images? Are the cells moving or do you have evidence that some of these changes in the phasor analysis don't result from jitter in the positions of the cells?
-
Here, we take a stack of 600 sequential qOBM images taken at 8 Hz (this frame rate was selected to capture metabolic activity within the cell)
Is the expectation here that metabolic changes will cause changes in the refactive index on a time scale of greater than 8 Hz? Is that a realistic time frame for metabolic changes? or are these other structural changes in response to activation?
-
