Abstract
Recently, Immunohistochemistry (IHC) using specific antibodies has been exploited to detect the localization of stem cells and cancer stem cells (CSCs). Typically, there is a very small population of stem cells in normal and cancerous tissues, making it difficult to investigate their cell type and localization in human specimens. Hence, for pathological detection of human specimens, IHC has certain advantages over immunofluorescence staining. In the present study, we established a method for double-staining cancer stem cell markers and other putative markers associated with human liver cancer. We have summarized and reviewed the methodology for IHC with doublestaining in human liver cancer tissues.
Highlights
Immunohistochemistry (IHC) is a useful technique for detection of proteins in cells and is applicable for diagnosis and research in both human cancer and normal tissues
For double-staining, ALDH1A1 and CD90, ALDH1A1 and BMI1, ALDH1A1 and epithelial cell adhesion molecule (EpCAM), and ALDH1A1 and Ki67 were used as pairs (Figure 1)
ALDH1A1 was first stained brown by DAB, while CD90, EpCAM, BMI1, and Ki67 were stained blue by alkaline phosphatase (AP)
Summary
Immunohistochemistry (IHC) is a useful technique for detection of proteins in cells and is applicable for diagnosis and research in both human cancer and normal tissues. IHC has several advantages over IF technique including: 1) capability of being repeatedly used in order to search for few positive cells, 2) no reduction in signal intensity, 3) provides superior morphological information, and 4) could be utilized to distinguish between normal and cancerous tissue. In cancerous tissues, double-staining IHC is practically suitable for observing stem cell and CSC markers [3,4]. Cancer tissues are hierarchically organized as they are composed of heterogeneous cell populations including a very small fraction of CSCs. Limited methods are available to purify CSCs owing to tumor cell heterogeneity and lack of specific stem cell markers. ALDH1A1, a major isoform of ALDH1, has been reported as a differentiation marker for the detection of HCC It could only detect, well, moderately, or poorly differentiated HCCs, rather than the CSC fraction by IHC. We reviewed and summarized a unique, modified doublestaining protocol used in our laboratory for years to detect stem cell markers in human formalin-fixed and paraffin-embedded liver cancer tissue specimens
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
