The BioGenex i6000® automated staining system, antibodies, and reagents were used by researchers to develop an ultra-rapid cytokeratin (CK) immunostaining protocol for Mohs micrographic surgery (MMS) which otherwise uses a lengthy time-consuming protocol.
Mohs micrographic surgery is a well-established surgical procedure for removing Basal Cell Carcinoma (BCC) and Squamous Cell Carcinoma (SCC). The technique often involves removing a thin layer of tissue one at a time and having the tissues H&E stained for marginal characteristics during the surgery rather than post-surgery. This ensures complete removal of the cancerous tissue with minimal noncancerous tissue loss to the patient. However, failure in detecting and differentiating the tumor cells from cancerous cells using H&E staining has led to a recurrence of tumors in about 1-2% in cases of BCC and 3.1% in SCC.
A team of researchers from the University of South Florida has chosen BioGenex products to develop a rapid protocol for frozen-section CK immunostaining of non-melanoma skin cancer (NMSK). CK immunostaining using pan-cytokeratin antibodies provides increased confidence in differentiating the epithelial and mesothelial cells from mesenchymal cells in normal and tumor tissues, especially for previously negative H&E tissues. NMSK show characteristics of over-expression of various classes of cytokeratins. The BioGenex AE1/AE3 – anti-CK antibody cocktail was chosen to target a full spectrum of keratin polypeptides, both acidic and basic to enhance the detection level. AE1 targets CK10, 14, 15, 16 and 19; AE3 targets CK1, 2, 3, 4, 5, 6 and 7. BioGenex’s citrate buffer was used along with heat-induced antigen retrieval method. This buffer has been proven to work with the most common and difficult-to-retrieve antigens and is compatible with most of BioGenex’s antibodies.
The frozen sections from 21 patients who underwent MMS were subjected to the ultra-rapid protocol using the BioGenex i6000® automated staining system. For all cases, samples were submitted for permanent (FFPE) CK staining and H&E staining. The rapid CK results were compared with permanent (FFPE) CK immunostaining which showed similar results, suggesting the accuracy of the ultra-rapid 19-minute protocol. The study results were published in the Dermatologic Surgery.
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