Comparison of flow cytometry and immunohistochemistry in non-radioisotopic murine lymph node assay using bromodeoxyuridine

Kyoung Mi Jung, Il Hong Bae, Bae Hwan Kim, Wang Ki Kim, Jin Ho Chung, Young Ho Park, Kyung Min Lim

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21 Scopus citations


Non-radioisotopic local lymph node assay (LLNA) employing 5-bromo-2′-deoxyuridine (BrdU) with flow cytometry (FACS) or immunohistochemistry (IHC) is gaining attention due to a regulatory issue of using radioisotope, 3H-thymidine, in vivo in traditional LLNA. In this study, to compare the performance of these non-radioisotopic endpoints, 7 chemicals with known sensitizing potencies were examined in LLNA. Mice were topically treated with chemicals or vehicle on both ears for 3 days. After intraperitoneal injection of BrdU, bilateral lymph nodes were isolated separately and undergone respectively, FACS or IHC to determine BrdU incorporated lymph node cells (LNCs). Weight and histology of treated ears were also examined to evaluate chemical-induced edema and irritation. Both FACS and IHC could successively identify the skin sensitizers from non-sensitizers. Comparison of FACS and IHC with traditional LLNA revealed that FACS has a higher sensitivity although both assays produced comparable sensitivity and performance to traditional LLNA. In conclusion, non-radioisotopic LLNA using FACS and IHC can successfully detect sensitizers with a good correlation to traditional LLNA. Notably, FACS showed almost equivalent sensitivity and accuracy to traditional LLNA.

Original languageEnglish
Pages (from-to)229-237
Number of pages9
JournalToxicology Letters
Issue number2
StatePublished - 1 Feb 2010

Bibliographical note

Funding Information:
This research was supported by a grant ( 09162KFDA557 ) from Korean Food & Drug Administration in 2009.


  • 5-Bromo-2′-deoxyuridine
  • Flow cytometry
  • Immunohistochemistry
  • Non-radioisotopic local lymph node assay


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