Overcoming fixation and permeabilization challenges in flow cytometry by optical barcoding and multi-pass acquisition.

ABSTRACT: The fixation and permeabilization of cells are essential for labeling intracellular biomarkers in flow cytometry. However, these chemical treatments often alter fragile targets, such as cell surface and fluorescent proteins (FPs), and can destroy chemically-sensitive fluorescent labels. This reduces measurement accuracy and introduces compromises into sample workflows, leading to losses in data quality. Here, we demonstrate a novel multi-pass flow cytometry approach to address this long-standing problem. Our technique utilizes individual cell barcoding with laser particles, enabling sequential analysis of the same cells with single-cell resolution maintained. Chemically-fragile protein markers and their fluorochrome conjugates are measured prior to destructive sample processing and adjoine

SUBMITTER: Fahlberg MD 

PROVIDER: S-EPMC12707249 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature