Fikar, P. | bioRxiv Science

bioRxiv2024

Enhancing reproducibility and decentralization in single cell research with biocytometry

Biomedicine today is experiencing a shift towards decentralized data collection, which promises enhanced reproducibility and collaboration across diverse laboratory environments. This inter-laboratory study evaluates the performance of biocytometry, a method utilizing engineered bioparticles for enumerating cells based on their surface antigen patterns. In a decentralized framework, spanning 78 assays conducted by 30 users across 12 distinct laboratories, biocytometry consistently demonstrated significant statistical power in discriminating numbers of target cells at varying concentrations as low as 1 cell per 100,000 background cells. User skill levels varied from expert to beginner capturing a range of proficiencies. Measurement was performed in a decentralized environment without any instrument cross-calibration or advanced user training outside of a basic instruction manual. The results affirm biocytometry to be a viable solution for immunophenotyping applications demanding sensitivity as well as scalability and reproducibility and paves the way for decentralized analysis of rare cells in heterogeneous samples.

cell biology↗

bioRxiv2024

Massively parallel identification of single-cell immunophenotypes

Translating insights from single-cell analysis into actionable indicators of health and disease requires large-scale confirmatory studies. We introduce biocytometry, a novel method utilizing engineered bioparticles for multiparametric immunophenotyping in suspension, enabling simultaneous measurement across thousands of assays with single-cell sensitivity and a wide dynamic range (1 to 1,000 target cells/sample). The technical validation of biocytometry revealed strong alignment with established technologies (mean bias = 0.25%, LoA = -1.83% to 2.33%) for low-sensitivity settings. Biocytometry excelled in high-sensitivity settings, consistently showcasing superior sensitivity and specificity (LoB = 0), irrespective of the sample type. By employing multiparametric target cell identification, we harnessed the homogeneous assay workflow to discern cell-specific apoptosis in mixed cell cultures. Potential applications include monitoring rare premalignant subpopulations in indications such as smoldering multiple myeloma (SMM), enhancing the detection of circulating tumor cells (CTCs), advancing pharmacokinetic assessments in chimeric antigen receptor (CAR) T-cell therapies, and improving the accuracy of minimal residual disease (MRD) evaluations. Additionally, the high throughput and cell-specific readout capabilities might provide substantial value in drug development, especially for the analysis of complex sample matrices, such as primary cell cultures and organoids.

synthetic biology↗