Imaging Cytometry represents a significant advancement in modern scientific research, merging the swift analytical power of cytometry with the detailed cellular insights provided by cutting−edge imaging technologies. Imaging Cytometry has evolved remarkably from its initial concept in late 1970s. This evolution overcame initial obstacles, such as developing sophisticated data acquisition and analysis software. In our session, we will explore the forefront of initial applications, showcasing a variety of advanced techniques from high-throughput, label-free cell imaging and multi−parametric analysis for precise protein identification. To enhance image quality and reduce artifacts for a better data exploration, advanced denoising methods have been developed utilizing advanced deep-learning image processing algorithms. Furthermore, the incorporation of artificial intelligence (AI) into Imaging Cytometry has significantly improved our ability to analyze cellular morphology, opening new paths in cell biology and disease research. Our discussion will also cover the challenges in Imaging Cytometry implementation and the intricacies of AI integration for data analysis, and the ongoing quest for improvements in sensitivity and specificity. The session aims to offer an in−depth look at Imaging Cytometry’s current capabilities, the essential tools for its application, the obstacles to be overcome, and the future enhancements that could further empower this technology in scientific research.

CMLE Credit: 1.0

Since 2022, the Indonesian Government has expanded the provision of leukemia and lymphoma immunophenotyping testing across the country from 9 sites to a projected 35 sites by 2025 to cover all the regional provincial hospitals. This increase in the number of labs doing this test required education of lab staff and development of assay standardization. To aid the education process, flow cytometry workshops have been conducted in Indonesia with the collaboration with the ISAC Live Education Task Force. These workshops included lectures and labs which covered the principles of flow cytometry, immunofluorescence techniques for flow cytometry, and its use in immunophenotyping of haematological malignancies. Development of standardized assays also plays a crucial role to be able to scale-up the capacity and provision of clinical services. A simplified 3-tube-8-color panel was designed, its performance verified, and then implemented in most of the testing sites after specialized training. In this scientific tutorial we will discuss how this was done and where we are in this process. We would also like to hear from any others who have undertaken a similar activity.

CMLE Credit: 1.0

This talk will be a light touch walk-through different approaches to label-free Quantitative Phase Imaging and how to extract and maximise the rich data. QPI is arguably the richest form of cytometry, with the ability to study the mass, volume, size, shape, granularity, and many phenotypic traits over time-lapse imaging without having to label the cells. There are multiple approaches to obtain QPIs including ptychography and digital holography. We will look at both approaches and the types of data that can be collected and scientific studies for which they are particularly useful. We will also look at the data analysis approaches that can make light work for the non-expert user.

CMLE Credit: 1.0

With the increase in panel sizes, more and more people are interested in computational cytometry tools to supplement their manual gating strategies. In this tutorial, I will give an overview of the typical computational cytometry pipeline we use in our lab. I will introduce several types of algorithms, what their strengths and caveats are and in which situations they might be applicable. The algorithms will be demonstrated both on a toy dataset, to showcase the principles and which will be made available for further exploration by the participants, as well as on concrete examples of studies we worked on in our lab. At the end, the audience should have a clear idea of some computational tools that might be worth trying on their own use cases.

CMLE Credit: 1.0

Instrument performance characterization and monitoring can help you identify developing problems in time to address them proactively. Being able to provide this type of data can also help you get the appropriate vendor-provided service more rapidly when the repair exceeds in-house abilities or resources. We’ll discuss setting up performance monitoring plans in an SRL environment and how to use them to inform troubleshooting. Approaches covered will include, but are not limited to: initial performance characterization, manufacturer and supplemental QCs, and the utility of Levy-Jennings plots. This session is geared toward leaders and staff of new or growing SRLs but welcomes all cytometrists looking to discuss the topic.

CMLE Credit: 1.0

In recent decades, the field of cytometry has witnessed a remarkable surge in the adoption of new fluorescent dyes, driving the widespread integration of large panels and spectral technologies into daily workflows. These developments have underscored the critical need to account for dye interactions within the reagent mix and during cell staining procedures. However, these interactions are often not sufficiently described, primarily due to the scarcity of unbiased technical reports, further compounded by the proprietary nature of reagent information. In this tutorial, we will first briefly examine the chemistry families of fluorescent dyes commonly employed in flow cytometry. We will then revisit established considerations regarding unwanted interactions between reagents, as well as between dyes and cells. This includes discussions on Fc receptor binding, cyanine- and cyanine-like dye interactions with myeloid cells, polymer dye aggregation, undesired FRET detection, and other pertinent factors. The primary emphasis of the tutorial will center on what the community regards as 'novel dyes' and their potential unwanted interactions with both reagent mix components and target cells. Furthermore, we will explore strategies for future-proofing panel designs to accommodate the addition of reagents, as well as addressing adding entities with varying brightness levels to the panels. These discussions will provide attendees with valuable insights into optimizing flow cytometry panel designs to effectively navigate the evolving landscape of cytometric analyses.

CMLE Credit: 1.0