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TECHNICAL REPORT Ki-67 Membranous Staining: Biologically Relevant or an Artifact of Multiplexed Immunofluorescent Staining Dan Wang, MSc,* Zhengyu Pang, PhD,w Gina M. Clarke, PhD,* Sharon Nofech-Mozes, MD,zy Kela Liu, MD,* Alison M. Y. Cheung, PhD,* Robert J. Filkins, PhD,w and Martin J. Yaffe, PhD*8 time due to the spectral overlap of fluorescent dyes. To Abstract: In the process of developing a multiplex of 8 common overcome these limitations, multiepitope ligand cartog- breast cancer biomarkers (Her2/neu, estrogen receptor, proges- raphy used photo-bleaching to create topological maps of + + 1,2 terone receptor, Ki-67, aldehyde dehydrogenase-1, Na K -AT- hundreds of proteins. At General Electric Company, a Pase, cytokeratin 8/18, and myosin smooth muscle) on a single novel multiplexed fluorescence microscopy method (MxIF) formalin-fixed paraffin-embedded slide using a sequential staining, employing chemical inactivation of fluorescent dyes after imaging, and dye bleaching technology developed by General imaging has been developed, and is currently commer- Electric Company, membranous Ki-67 staining was observed and cialized as MultiOmyx at Clarient (a GE HealthCare colocalized with Her2/neu staining. Using immunohistochemistry Company). This allows the same fluorophores to be reused as gold standards, we discovered that membranous Ki-67 was an on different dye-conjugated primary antibodies in an
Applied Immunohistochemistry & Molecular Morphology – Wolters Kluwer Health
Published: Aug 1, 2015
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