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Boushehri, S. S.

Publications and source records attributed to Boushehri, S. S..

2 recordsLinked to original sources

PXPermute: Unveiling staining importance in multichannel fluorescence microscopy

Imaging Flow Cytometry (IFC) enables rapid acquisition of thousands of single-cell images per second, capturing information from multiple fluorescent channels. However, the conventional process of staining cells with fluorescently labeled conjugated antibodies for IFC analysis is labor-intensive, costly, and potentially detrimental to cell viability. To streamline experimental workflows and reduce expenses, it is imperative to identify the most relevant channels for downstream analysis. In this study, we present PXPermute, a user-friendly and powerful method that assesses the significance of IFC channels for a given task, such as cell profiling. Our approach evaluates channel importance by permuting pixel values within each channel and analyzing the resulting impact on the performance of machine learning or deep learning models. Through rigorous evaluation on three multi-channel IFC image datasets, we demonstrate the superiority of PXPermute in accurately identifying the most informative channels, aligning with established biological knowledge. To facilitate systematic investigations of channel importance and aid biologists in optimizing their experimental designs, we have released PXPermute as an easy-to-use open-source Python package.

bioinformatics↗

Spatial proteomics in neurons at single-protein resolution

To fully understand biological processes and functions, it is necessary to reveal the molecular heterogeneity of cells and even subcellular assemblies by gaining access to the location and interaction of all biomolecules. The study of protein arrangements has seen significant advancements through super-resolution microscopy, but such methods are still far from reaching the multiplexing capacity of spatial proteomics. Here, we introduce Secondary label-based Unlimited Multiplexed DNA-PAINT (SUM-PAINT), a high-throughput imaging method capable of achieving virtually unlimited multiplexing at better than 15 nm spatial resolution. Using SUM-PAINT, we generated the most extensive multiprotein dataset to date at single-protein spatial resolution, comprising up to 30 distinct protein targets in parallel and adapted omics-inspired analysis workflows to explore these feature-rich datasets. Remarkably, by examining the multiplexed protein content of almost 900 individual synapses at single-protein resolution, we revealed the complexity of synaptic heterogeneity, ultimately leading to the discovery of a new synapse type. This work provides not only a feature-rich resource for researchers, but also an integrated data acquisition and analysis workflow for comprehensive spatial proteomics at single-protein resolution, paving the way for Localizomics.

neuroscience↗