Macrophage Immunology Assays on Livecyte
Livecyte combines high contrast label-free imaging with correlative fluorescence and automatic cell tracking algorithms to provide a complete solution for quantifying phagocyte behaviour down to the single-cell level. This enables users to…
- Automatically quantify macrophage phenotypic behaviour label-free to measure immune response
- Quantification to the single-cell level gives a more accurate measure of phagocytosis than standard population-level analyses
- Increased accuracy reveals subtle behaviour changes from how phagocyte activity changes over time to saturation
Quantifying Phenotypic Changes to Measure Immune Response
Macrophages are known to change their behaviour or state in response to specific functional initiatives. Mediators Interferon γ (IFNγ) and bacterial lipolysaccaride (LPS) promote a more active pro-inflammatory M1 phenotype, but measuring this response is challenging.
The quantitative phase images produced by Livecyte are ideal for single-cell segmentation and tracking enabling automatic quantification of phenotypic behaviour such as cell morphology, motility and proliferation.
This allows macrophage immune response to be measured in an assay relevant format by the quantified change in phenotypic behaviour.
Single-cell Phagocytosis Measurement
A key function in macrophage immune response is the phagocytosis of foreign bodies or pathogens. Traditionally this is quantified by measuring the fluorescence expression of an appropriate biomarker across a whole image. However this population-level approach over simplifies complex single-cell level mechanics and population heterogeneity.
Livecyte quantifies correlative fluorescence expression down to the single-cell level eliminating confounding effects from cell proliferation and preserving the heterogeneity of the response.
On Livecyte, fluorescence imaging can be performed intermittently as individual cells are tracked using the QPI modality. This substantially reduces phototoxicity for the assay.
Reveal How Phagocyte Activity Changes Over Time
Efferocytosis, the phagocytosis of apoptotic cells, is critical for the maintenance of tissue health and reducing inflammation.
Livecyte can combine the quantification of phagocytosis, and the measurement of macrophage phenotypic behaviour in a single experiment which can lead to insights not possible from either assay in isolation.
For example in AN019 we correlate phagocyte activity with changes in macrophage motile behaviour and label-free growth and proliferation identifying changes in individual phagocyte activity over time and potential phagocyte saturation.
One professor has been using the Livecyte on a daily basis and his work flow has become so streamlined that he has managed to get a few months’ worth of work done in just one.
Greg Perry, St George’s University of London
