Comparison of cell fixation methods of induced sputum specimens: an immunocytochemical analysis.
Induced sputum (IS) is a non-invasive method to evaluate airway inflammation. Various techniques are used to fix IS cells but their respective value has never been compared. We aimed to determine the best IS cell fixation technique for cellular markers staining. Cells were fixed using four methods: 1) periodate-paraformaldehyde-lysine (PLP)-sucrose, 2) paraformaldehyde 4% on slide and 3) in solution and 4) acetone-methanol. Analysis was based on percentage of positive cells compared to total cell counts stained by hematoxylin and quality of staining. Using PLP-sucrose resulted in a higher percentage of positive cells for CD3 and a better quality of staining. Acetone-methanol showed a lower percentage of positive cells for CD68 and a poor quality. PLP-sucrose gives the best results for the preservation of the studied cell markers and acetone-methanol the worst.
St-Laurent
Julie
J
Unité de recherche en pneumologie, Centre de recherche de l'Hôpital Laval, Institut universitaire de cardiologie et de pneumologie, Québec, Québec, Canada.
Boulay
Marie-Eve
ME
Prince
Philippe
P
Bissonnette
Elyse
E
Boulet
Louis-Philippe
LP
D000328:Adult; D015703:Antigens, CD / Q000378:metabolism; D015214:Antigens, Differentiation, Myelomonocytic / Q000378:metabolism; D017252:CD3 Complex / Q000378:metabolism; D047150:Eosinophil Cationic Protein / Q000378:metabolism; D005260:Female; D005404:Fixatives; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D019272:Leukocyte Elastase / Q000378:metabolism; D007962:Leukocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D008297:Male; D013183:Sputum / Q000166:cytology* / Q000276:immunology*; D016707:Tissue Fixation / Q000379:methods*; D000097382:CD68 Molecule
D015703:Antigens, CD; D015214:Antigens, Differentiation, Myelomonocytic; D017252:CD3 Complex; D047150:Eosinophil Cationic Protein; D005404:Fixatives; D019272:Leukocyte Elastase; C067980:CD68 antigen, human; D000097382:CD68 Molecule; C479220:RNASE3 protein, human