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Comparison of histochemical methods for murine eosinophil detection in a RSV vaccine‐enhanced inflammation model
Author(s) -
Griffin Michelle Ann,
Castilow Elaine M,
Varga Steven M,
Meyerholz David K
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.571.6
Subject(s) - staining , eosinophil , stain , sirius red , pathology , h&e stain , inflammation , biology , medicine , microbiology and biotechnology , chemistry , immunology , asthma
Five histochemical methods previously reported for eosinophil detection were quantitatively and qualitatively compared in an established murine RSV vaccine‐enhanced inflammation model. Nonspecific neutrophil staining was evaluated in tissue sections of neutrophilic lesions and bone marrow from respective animals. Eosinophils had granular red to orange‐red cytoplasmic staining, depending on the method, whereas neutrophils had a more homogenous cytoplasmic pattern. Nonspecific background staining of similar coloration was variably seen in arterial walls and erythrocytes. Astra Blue/Vital New Red (AB/VNR), Congo Red, Luna, Modified Hematoxylin & Eosin (Mod HE), and Sirius Red techniques were all effective (p<0.001) in detecting increased eosinophil recruitment compared to controls; however, differences in absolute eosinophil quantification were seen. AB/VNR had the best specificity for differentiating eosinophils and neutrophils, but had a significantly reduced ability to enumerate eosinophils. The Luna stain had excessive non specific staining of tissues and a reduced enumeration of infiltrating eosinophils making it suboptimal. For multiple parameters such as eosinophil detection, specificity, and contrast with background tissues, the Sirius Red followed by Congo Red and Mod HE methods were useful, each with their own staining qualities. Supported by NIH and UI Department of Pathology.