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Updated: Aug 9, 2026
Fluorescence-mediated Tomography for the Detection and Quantification of Macrophage-related Murine Intestinal Inflammation
Published on: December 15, 2017
A fluorescence method for the visualization of inflammatory cells in intestinal mucosa
1Department of Pharmacology, Pharmacia AB, Uppsala, Sweden.
Insights
Unlabeled fluorescein isothiocyanate (FITC) staining offers a rapid method for identifying phagocytic cells like neutrophils and macrophages in tissue samples. This technique shows high correlation with established methods, making it a convenient diagnostic tool.
Area of Science:
- Histopathology
- Immunofluorescence
- Cell Biology
Background:
- Accurate identification of phagocytic cells is crucial in diagnosing various pathological conditions.
- Traditional methods for visualizing these cells can be time-consuming and complex.
Purpose of the Study:
- To evaluate the efficacy of unlabeled fluorescein isothiocyanate (FITC) staining for identifying phagocytic cells in histological specimens.
- To compare the novel FITC staining method with established immunofluorescence and histochemical techniques.
Main Methods:
- Paraffin and cryostat sections from rat, rabbit, and human tissues were stained with unlabeled FITC.
- FITC staining was compared against FITC-conjugated anti-neutrophil antibodies, endogenous peroxidase staining, and FITC-conjugated Lotus tetragonolobus lectin staining.
- Receptor inhibition studies were performed using unconjugated antibodies and L-fucose.
Main Results:
- Unlabeled FITC staining effectively visualized fluorescent leukocytes with granular patterns in light microscopy.
- FITC staining demonstrated strong correlations with all three comparative methods (r values of 0.97-0.98, P < 0.001).
- Pre-incubation with specific inhibitors abolished staining, confirming the specificity of the FITC reaction.
Conclusions:
- Unlabeled FITC staining is a quick, convenient, and reliable method for estimating phagocytic cells (polymorphonuclear cells and macrophages) in histological samples.
- This method offers a valuable alternative for routine histopathological analysis.
Abstract:
Treatment of paraffin or cryostat sections from rat, rabbit and man, with fluorescein isothiocyanate (FITC) in buffer (1 microgram/l), resulted in visualization of highly fluorescent leukocytes with a granular staining in light microscopy. FITC staining of paraffin-embedded colonic tissue was compared with three other staining methods, i.e., (1) FITC-conjugated anti-neutrophil antibodies, (2) endogenous peroxidase with 3,3-diaminobenzidine, and (3) FITC-conjugated Lotus tetragonolobus lectin. Good correlations between FITC staining and the three other methods were found (r:0.97, 0.98 and 0.98; P less than 0.001). Inhibition of receptors for the FITC conjugates prior staining, by preincubation with unconjugated anti-neutrophil antibodies or L-fucose, abolished the staining with antibodies and lectin. This study shows that staining with unlabeled FITC represents a quick and convenient method for the estimation of phagocytic cells (polymorphonuclear cells and macrophages) in histological specimens.
