Comparison of Methods for Analyzing Human Adipose Tissue Macrophage Content
Maria Morgan-Bathke1, Debra Harteneck1, Philippa Jaeger1
1Endocrine Research Unit, Mayo Clinic, Rochester, Minnesota, USA.
Obesity (Silver Spring, Md.)
|October 7, 2017
Summary
This study compared methods for measuring adipose tissue macrophages (ATM), finding that the AMCounter software improves reproducible and efficient immunohistochemistry (IHC) measurements. Different techniques offer distinct insights into adipose inflammation.
Area of Science:
- Immunology
- Metabolic Health
- Cell Biology
Background:
- Adipose tissue macrophages (ATM) are linked to inflammation, obesity, and metabolic dysfunction.
- Accurate quantification of ATM is crucial for understanding these conditions.
Purpose of the Study:
- To compare the efficacy of flow cytometry, immunohistochemistry (IHC), and RT-PCR in measuring human ATM.
- To evaluate a new software (AMCounter) for enhancing IHC-based ATM measurements.
Main Methods:
- Human ATM were measured using flow cytometry, IHC, and RT-PCR.
- The AMCounter software was assessed for its impact on IHC measurements.
- Reproducibility and efficiency of each method were compared.
Main Results:
- IHC demonstrated good reproducibility for ATM markers (CD68, CD14, CD206).
- AMCounter improved IHC interreader agreement and efficiency.
- Flow cytometry showed acceptable reproducibility for ATM percentages but not per tissue gram.
- IHC and flow cytometry measures of ATM per gram were discordant and uncorrelated.
- RT-PCR showed significant correlations with IHC and flow cytometry measures.
Conclusions:
- AMCounter software enhances the reproducibility and efficiency of IHC-based ATM quantification.
- Flow cytometry, IHC, and RT-PCR provide complementary, rather than interchangeable, data on adipose tissue inflammation.
