Investigating the murine hepatic immune composition in diet-induced obesity using OMIP-104: Transferring an existing OMIP panel onto the CytoFLEX mosaic Spectral Detection Module
Executive Summary
Obesity-induced chronic inflammation (meta-inflammation) results from an altered immune response in metabolic organs and contributes to the development of fatty liver disease and type 2 diabetes. To dissect these complex immune dynamics, OMIP-104 — a 30-color spectral flow cytometry panel — was developed to characterize resident and recruited macrophage subsets, macrophage and dendritic cell activation states, and major leukocyte populations in murine metabolic tissues (liver and white adipose tissue).
This application note demonstrates the transfer of OMIP-104 onto the CytoFLEX LX Flow Cytometer equipped with the CytoFLEX mosaic 88 Spectral Detection Module, conducted without altering the published panel.
Key Highlights & Findings
- Panel Transferability: Successfully identified all key hepatic leukocyte populations and their subsets — including Kupffer cells (resident, monocyte-derived, and pre-monocyte-derived), dendritic cells (cDC1s and cDC2), T cells, NK cells, B cells, and monocyte subsets — without altering the published panel.
- Multi-Signature Autofluorescence (AF) Management: Extracting and subtracting four unique autofluorescent signatures identified from unstained samples reduced unmixing errors and improved cell population resolution compared with using a single autofluorescent signature.
- Minimal Panel Complexity: The panel achieved a complexity score of 11.0, with no fluorochrome combination exceeding a similarity index of 0.84, indicating a minimal risk of excessive spread errors.
- Flexible Unmixing Algorithms: Both the commonly used Least Squares Method (LSM) and a proprietary modification of the Poisson algorithm performed similarly in defining and resolving cell populations for this panel, as shown on opt-SNE visualizations.
- Expansion Potential: Because no fluorochromes designed for the 808 nm near-infrared (IR) laser were included, this laser range offers a straightforward opportunity to expand the panel or replace fluorochromes that result in spread errors.
For Research Use Only. Not for use in diagnostic procedures.
