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Label Free Detection of Subtle Morphological Differences in Immune Cells

Sensitive and high-throughput technologies to assess subtle morphological differences in cells are a major need for high-throughput drug screening and disease profiling research. However, conventional technologies like flow cytometry and microscopy to assess morphology have limitations. Microscopy is slow and a terminal assay for cells while flow cytometry only provides crude physical measures, like size and granularity, by detecting forward (FSC) and side (SSC) scattered light, respectively. By these measures, subtle morphological differences between cells cannot be detected. However, the data-rich profiles generated by Ghost Cytometry enable detection and resolution of ultra-fine differences in cell morphology. In this proof-of-concept study we use the VisionSort platform, powered by Ghost Cytometry, to […]

Label-Free High Throughput CRISPR-Based Screening for Morphological Phenotypes in Flow

The development of CRISPR-Cas9 genome editing technology has led to the emergence of a new generation of novel life sciences applications. In drug discovery, researchers have harnessed the precision of selective gene knockouts by CRISPR to enable genome-wide drug screening. By mapping genotypes to phenotypes, CRISPR-based phenotypic screens can enable a better understanding of drug mechanism of actions (MOAs) and identification of novel druggable targets. However, current phenotypic CRISPR screening approaches rely heavily on microscopic imaging of target phenotypes, a process that imposes throughput limitations and restricts screening to only a handful of simple phenotypes based on binary fluorescence signals. To fully realize the potential of CRISPRbased phenotypic screening, here […]

High Throughput CRISPR-Based Phenotypic Screening in Flow for Complex Intracellular Phenotypes

The development of CRISPR-Cas9 genome editing technology has led to the emergence of a new generation of novel life sciences applications1. In drug discovery, researchers have harnessed the precision of selective gene knockouts by CRISPR to enable genome-wide drug screening. By mapping genotypes to phenotypes, CRISPR-based phenotypic screens can enable a better understanding of drug mechanism of actions (MOAs) and identification of novel druggable targets. However, current phenotypic CRISPR screening approaches rely heavily on microscopic imaging of target phenotypes, a process that imposes throughput limitations and restricts screening to only a handful of simple phenotypes by binary fluorescence signals. To fully realize the potential of CRISPR-based phenotypic screening, here show […]

Intracellular Phenotyping with Enhanced Spatial Resolution in Flow

Many cellular processes and functions involve phenotypic changes or biomolecular interactions at the intracellular level. Researchers can exploit knowledge of these phenotypic changes to better elucidate basic cellular biology, profile disease pathology, or identify new drug targets. Classically, cellular phenotypes have been assessed using high-resolution, imagebased microscopy approaches. However, microscopy-based approaches are slow, manual, and time consuming. Recently, phenotypic assessment using flow cytometry has been developed, offering substantially higher throughput. However, conventional flow cytometers only collect data on the average intensity of light signals and can not deeply analyze intracellular phenotypes with a high degree of spatial resolution. In addition, imaging flow cytometers are unable to sort cells based on […]

B cell Plasma Cell Differentiation

B cells are critical for a functional immune system. Activation of B cells results in a series of biochemical and cellular changes that convert naïve B cells to plasma cells. Differentiated plasma cells help the body to defend against foreign entities through the production of antibodies. This antigen-specific antibody production by plasma cells has also been exploited for the development of targeted therapeutics for cancer and other immune disorders. While B cells play a protective role in the adaptive immune system, dysfunctional B cells can lead to life threatening immunological disorders. Due to the importance of B cells for human health, studying the process of B-cell activation and plasma cell […]