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Title: NOVEL STAINING METHOD TO ENHANCE VISUALIZATION OF THE CHICKEN PEYER'S PATCH IN SITU

Author
item Vaughn, Lara
item Holt, Peter
item Moore, Randle

Submitted to: Poultry Science Association Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 3/16/2005
Publication Date: 7/31/2005
Citation: Vaughn, L.E., Holt, P.S., Moore, R.W. 2005. Novel staining method to enhance visualization of the chicken peyer's patch in situ. Poultry Science Association Meeting Abstract.

Interpretive Summary:

Technical Abstract: The chicken ileal Peyer's patches (Pp) are classified as secondary gut-associated lymphoid tissue of the mucosal immune system. Young birds may possess multiple Peyer's patches, yet as the bird ages these Pp sites tend to regress in number. Also, Peyer's patches of aging chickens are often difficult to observe grossly. There is one Pp located anterior to the ileocecal junction which appears to be consistently present in a range of age groups. This single Pp may serve as an important tissue site for monitoring inflammatory & immunological responses of the host against enteric pathogens. However, a simple technique to enhance visualization of the Pp is lacking. Therefore, we designed a novel staining method that is quick, easy & accurate to aid in gross identification & recovery of the chicken Pp. Segments of lower GI tract, jejunoileal junction extending distally to colon, were excised from SPF white leghorn spent hens & commercial broilers 4-6 wks of age. GI segments were flushed with phosphate buffered saline (PBS) to remove ingesta & cellular debris. Eosin-Y & a modified-Crystal Violet were then infused into the GI tract segments in a sequential fashion. First, Eosin-Y diluted in PBS was infused into the GI segment via 30cc syringe with 16 gauge, 1 inch needle until the GI segment was fully distended. The Eosin-Y stain was allowed to permeate for 1 minute then gently extruded. Second, modified-Crystal Violet diluted in PBS was infused into the GI segment. Upon addition of Crystal Violet, the lymphoid tissue area became apparent at the GI serosal surface. The distal ileal Pp appeared as a pale whitish-pink ovoid area with surrounding gut tissue stained light purple. The exact site of the Pp lymphoid tissue could be delineated with the unaided eye from both serosal & mucosal surfaces and then properly excised. Tissue was verified by histology. The novel Eosin-Y + Crystal Violet staining technique promotes rapid identification of & accurate recovery of chicken Pp lymphoid tissue from fresh tissue specimens. The method renders specific Pp lymphoid tissue suitable for use in various research applications. KEYWORDS Peyer's patch Eosin-Y Crystal Violet