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A simple method to demonstrate normal and metaplastic Paneth cells in tissue sections
1Gastrointestinal and Liver Pathology Research Laboratory, Karolinska Institute and Hospital, 171 76 Stockholm, Sweden. Carlos.Rubio@onkpat.ki.se
Abstract:
Using hematoxylin and eosin (H&E)-stained sections, Paneth cells are identified by the presence of eosinophilic granules. Several other methods have been proposed to stain Paneth cells. Recently, we noticed that Paneth cells were autofluorescent in H&E sections. To assess the number of Paneth cells by this method, 30 surgical specimens from the small bowel and 10 from the colon with IBD were investigated. Consecutive sections were stained with H&E and immunostained with lysozyme (Lz). Paneth cell counting was done in 20 well-oriented, vertically cut crypts per-case using alternatively transmitted light (TL) and incident-light fluorescence (ILF). When H&E and Lz sections were observed with TL, the number of Paneth cells was easily assessed in some crypts, but in crypts having numerous Paneth cells, their exact number was ascertainable. On the other hand, using IFL, the number of H&E-stained Paneth cells-crypt was easily assessed, even in crypts with Paneth cell clustering. Using H&E-ILF, a mean of 3.9 (range 3-8) Paneth cells-pe-crypt was found in the normal small bowel and a mean of 3.3 (range 1-14) in the colon with IBD. ILF provided the adequate visual conditions--i.e. a dark, non-autofluorescent background--for assessing the actual number of normal and metaplastic Paneth cells, even when arranged in tight cell groups. The present method will improve the enumeration of the actual number of normal and metaplastic Paneth cells both in experimental research and in clinical trials.
Insights
Autofluorescence in hematoxylin and eosin (H&E) stained sections allows for accurate Paneth cell counting. Incident-light fluorescence (ILF) improves enumeration of Paneth cells, especially in inflamed tissues.
Area of Science:
- Histopathology and Cell Biology
- Gastroenterology and Digestive Diseases
Background:
- Paneth cells are typically identified by eosinophilic granules in hematoxylin and eosin (H&E) stained sections.
- Accurate enumeration of Paneth cells can be challenging, particularly in cases with cell clustering or inflammation.
- Paneth cells exhibit autofluorescence under specific lighting conditions.
Purpose of the Study:
- To evaluate the utility of Paneth cell autofluorescence in H&E stained sections for accurate cell counting.
- To compare the effectiveness of incident-light fluorescence (ILF) versus transmitted light (TL) for Paneth cell enumeration.
Main Methods:
- Surgical specimens from the small bowel (n=30) and colon with inflammatory bowel disease (IBD) (n=10) were analyzed.
- Consecutive sections were stained with H&E and immunostained for lysozyme (Lz).
- Paneth cell counts were performed in 20 crypts per case using both TL and ILF on H&E sections.
Main Results:
- Incident-light fluorescence (ILF) enabled accurate assessment of Paneth cells, even in crypts with significant cell clustering.
- Transmitted light (TL) provided less precise counts in areas with numerous or clustered Paneth cells.
- Mean Paneth cell counts were 3.9 cells/crypt in normal small bowel and 3.3 cells/crypt in IBD colon using H&E-ILF.
Conclusions:
- Autofluorescence observed with ILF on H&E stained sections offers a reliable method for enumerating Paneth cells.
- This technique enhances visualization and improves the accuracy of Paneth cell quantification in both normal and metaplastic states.
- The H&E-ILF method is valuable for research and clinical trials requiring precise Paneth cell counts.
