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Differential interference contrast microscopy combined with immunofluorescence: a new method to phenotype eosinophils
F L Jahnsen1, G Haraldsen, J Rugtveit
1Laboratory for Immunohistochemistry and Immunopathology (LIIPAT), University of Oslo, National Hospital, Norway.
Insights
Differential interference contrast (DIC) imaging offers a reliable method for identifying eosinophils in tissue samples. This technique, combined with immunofluorescence, also enables detailed phenotyping of these cells.
Area of Science:
- Immunology
- Cell Biology
- Histopathology
Background:
- Eosinophil surface receptors vary with activation state in vitro.
- Current immunohistochemical studies often focus on eosinophil counts and granule protein deposits.
- Limitations exist in accurately identifying eosinophils in situ using traditional methods.
Purpose of the Study:
- To evaluate differential interference contrast (DIC) imaging for eosinophil identification in cryo-sections and cytospins.
- To compare the reliability of DIC imaging with immunohistochemical labeling for eosinophil detection.
- To explore the utility of DIC imaging combined with immunofluorescence for eosinophil phenotyping.
Main Methods:
- Differential interference contrast (DIC) imaging of cryo-sections and cytospins.
- Immunohistochemical labeling of eosinophil granule proteins.
- Three-color immunofluorescence microscopy.
- Combined DIC imaging and immunofluorescence microscopy.
Main Results:
- Eosinophils exhibit a distinct granular appearance under DIC imaging.
- DIC imaging proved more reliable for in situ eosinophil identification than immunohistochemistry.
- Combined DIC and immunofluorescence microscopy allowed for detailed three-color phenotyping of eosinophils.
Conclusions:
- DIC imaging is a superior method for identifying eosinophils in tissue samples.
- This technique enhances the ability to study eosinophil presence and characteristics in disease contexts.
- Combined DIC and immunofluorescence microscopy offers a powerful tool for eosinophil research.
Abstract:
Several surface receptors are expressed on eosinophils in vitro depending on the state of cellular activation, but immunohistochemical studies of eosinophil-related diseases have mostly focused on the number of infiltrating eosinophils as well as extracellular deposits of eosinophil granule proteins. The present investigation showed that eosinophils display a characteristic granular appearance in cryo-sections and cytospins by differential interference contrast (DIC) imaging. This approach appeared to be more reliable for identification of these cells in situ than immunohistochemical labelling of eosinophil granule proteins. Moreover, combined with immunofluorescence microscopy DIC imaging facilitated three-colour immunofluorescence phenotyping of eosinophils.
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