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The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
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A Mass Cytometry-Based Blood Cell Phenotyping Workflow Enabling Inclusion of Resource-Limited and Rural Sites in

Natalie J Smith1, Michael Cohen2, Lauren Tracey2

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A new mass cytometry workflow simplifies immune phenotyping for remote Australian communities. This approach ensures high-quality data for disease research, addressing health inequities in underserved populations.

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Area of Science:

  • Immunology
  • Biomedical Research
  • Clinical Trials

Background:

  • Comprehensive immune phenotyping is crucial for disease detection and treatment, but flow cytometry faces logistical challenges, especially in remote settings.
  • Remote and rural communities globally, including Australia, are underrepresented in research, leading to health inequities and poorer disease prognosis.
  • Existing methods require specialized equipment and stable reagent conditions, hindering studies in resource-limited areas.

Purpose of the Study:

  • To develop and validate a simplified mass cytometry workflow for high-parameter immune phenotyping in remote and rural Australian communities.
  • To enable the inclusion of underrepresented populations in biomedical research by overcoming logistical barriers.
  • To ensure the generation of high-quality immune profiling data from diverse geographical locations.

Main Methods:

  • Developed a mass cytometry workflow using dry-format, heavy-metal conjugated antibodies for 38 surface targets on whole blood (WB) samples.
  • Implemented cryopreservation of stained WB without centrifugation, simplifying sample preparation for resource-limited settings.
  • Centralized processing involved neutrophil removal, 6-channel barcoding, pooling, and staining for 8 intracellular targets, achieving 52-plex coverage.

Main Results:

  • Demonstrated workflow feasibility by characterizing immune and myeloid cell populations in cancer patients from three Australian sites, including a regional hospital.
  • Confirmed consistency in immune phenotype between the simplified WB approach and traditional peripheral blood mononuclear cell (PBMC) preparations.
  • Successfully included cancer patients from regional Australia in a multi-centre, high-parameter immune phenotyping study for the first time.

Conclusions:

  • The simplified mass cytometry workflow is compatible with resource-limited environments and conserves data quality.
  • This approach facilitates high-dimensional immune phenotyping in remote settings, enhancing research inclusivity.
  • The study addresses health inequities by enabling robust immune monitoring in underrepresented populations.