October 9 - 12, 2012
Building 10 & Natcher Conference Center
- General Schedule of Events
- Opening Plenary Session
- Concurrent Symposia Sessions
- Poster Sessions
- National Graduate Student Research Conference
- FARE Award Ceremony
- Special Exhibits on Resources for Intramural Research
- TSA Research Festival Exhibit Show
- Clinical Center Tours
- Research Festival Committees
- Past Research Festivals
Characterizing and diminishing autofluorescence in formalin-fixed paraffin-embedded human respiratory tissue
| Friday, November 08, 2013 — Poster Session IV | |||
|---|---|---|---|
2:00 p.m. – 4:00 p.m. |
FAES Academic Center (Upper-Level Terrace) |
NIAID |
VIROL-2 |
Authors
- AS Davis
- A Richter
- S Becker
- JE Moyer
- A Sandouk
- O Schwartz
- JK Taubenberger
Abstract
Tissue autofluorescence frequently hampers visualization of immunofluorescent markers in formalin-fixed paraffin-embedded tissues tissues. Focused specifically on human respiratory tissues, we sought to identify endogenous elements contributing to autofluorescence through multi-photon microscopic separation of elastin from collagen with immunofluorescent labeling for validation. We then assessed nine treatments reported to have efficacy in reducing autofluorescence in other tissue types. The three most efficacious were eriochrome black T, Sudan black and sodium borohydride as measured using white light laser confocal multi-lambda analysis. We also assessed the impact of steam antigen retrieval and serum application on human tracheal tissue autofluorescence. Functionally fitting this multi-lambda data to 2-dimensional Guassian surfaces revealed that steam antigen retrieval and serum application contribute minimally to autofluorescence and that the three treatments are disparately efficacious. Further, eriochrome black T when applied together with sodium borohydride showed an additive effect on reducing autofluorescence while combining Sudan black with sodium borohydride yielded no additional benefit. Together these studies provide a set of guidelines for diminishing autofluorescence in formalin-fixed paraffin-embedded human respiratory tissue. Additionally, the characterization techniques, including the mathematical model, should be transferable to similar questions in other tissue types.
