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The universal detection of antigens from one skin biopsy specimen
Haike M J van der Velden1, Peter C M van de Kerkhof, Marcel C Pasch
1Department of Dermatology, Radboud University Nijmegen Medical Centre, Nijmegen, the Netherlands. H.vanderVelden@derma.ucmn.nl
Insights
This study introduces a new method for immunohistochemistry, allowing detection of various antigens from a single skin biopsy. This technique eliminates the need for two separate biopsies, streamlining diagnostic processes in dermatology.
Area of Science:
- Dermatology
- Histopathology
- Immunohistochemistry
Background:
- Immunohistochemistry is crucial in dermatology but often requires two biopsies due to antigen preservation limitations.
- Certain antigens are only detectable on frozen sections, while others require formalin-fixed paraffin-embedded sections.
Purpose of the Study:
- To develop a universal detection technique for multiple antigens from a single biopsy specimen.
- To overcome the limitations of current immunohistochemistry methods in dermatology.
Main Methods:
- Single skin biopsies from psoriasis patients were processed for both frozen and paraffin-embedded sections.
- Frozen tissue was converted to paraffin-embedded sections by refixation in formalin.
- Various antigen retrieval techniques were applied, and differential expression of keratin 10, keratin 15, CD3, CD26, and human beta defensin-2 (HBD-2) was examined.
Main Results:
- Keratin 10 and 15 were successfully stained on both frozen and paraffin-embedded sections.
- CD3 and CD26 were only detectable on frozen sections.
- Human beta defensin-2 (HBD-2) was exclusively detected on paraffin-embedded sections.
Conclusions:
- A straightforward technique was developed to obtain both frozen and paraffin-embedded sections from a single biopsy.
- This method simplifies antigen detection in dermatological diagnostics, reducing the need for multiple biopsies.
Background:
Immunohistochemistry is an important tool in dermatology but is limited. Certain antigens can only be preserved in formalin-fixed paraffin-embedded sections, while others can only be detected on frozen sections, resulting in situations where two biopsies are needed. We aimed to develop a technique for universal detection of different antigens out of just one biopsy specimen.
Methods:
Single biopsies were obtained from lesional skin of patients with psoriasis. Standard sample procedures for frozen and paraffin-embedded sections were used. To convert frozen tissue into paraffin-embedded sections, the biopsy specimen was disposed of the embedding medium and subsequently fixed in 10% neutral buffered formalin. We applied various antigen retrieval techniques with alkaline solutions. The differential expression of keratin 10, keratin 15, CD3, CD26 and human beta defensin-2 (HBD-2) was examined using immunohistochemical staining.
Results:
We showed that keratin 10 and 15 can be stained on both frozen and paraffin-embedded sections. Staining of paraffin-embedded sections required unmasking with trypsin and Tris-buffered saline Tween solution, respectively. CD3 and CD26 can only be detected on frozen sections, while HBD-2 can only be detected on paraffin-embedded sections.
Conclusion:
We have described a straightforward technique that gives us the opportunity to use just one biopsy specimen to obtain frozen sections as well as paraffin-embedded sections.
