title: Enzyme catalysis enhanced dark-field imaging as a novel immunohistochemical method.
abstract: Conventional immunohistochemistry is limited to subjective judgment based on human experience and thus it is clinically required to develop a quantitative immunohistochemical detection. 3,3'-Diaminobenzidin (DAB) aggregates, a type of staining product formed by conventional immunohistochemistry, were found to have a special optical property of dark-field imaging for the first time, and the mechanism was explored. On this basis, a novel immunohistochemical method based on dark-field imaging for detecting HER2 overexpressed in breast cancer was established, and the quantitative analysis standard and relevant software for measuring the scattering intensity was developed. In order to achieve a more sensitive detection, the HRP (horseradish peroxidase)-labeled secondary antibodies conjugated gold nanoparticles were constructed as nanoprobes to load more HRP enzymes, resulting in an enhanced DAB deposition as a dark-field label. Simultaneously, gold nanoparticles also act as a synergistically enhanced agent due to their mimicry of enzyme catalysis and dark-field scattering properties.
authors.affiliation: State Key Laboratory of Bioelectronics, Jiangsu Key Laboratory for Biomaterials and Devices, School of Biological Science and Medical Engineering & Collaborative Innovation Center of Suzhou Nano Science and Technology, Southeast University, Nanjing 210096, P. R. China. zhangyu@seu.edu.cn guning@seu.edu.cn.
authors.forename: Lin
authors.initials: L
authors.lastname: Fan
authors.forename: Yanyan
authors.initials: Y
authors.lastname: Tian
authors.forename: Rong
authors.initials: R
authors.lastname: Yin
authors.forename: Doudou
authors.initials: D
authors.lastname: Lou
authors.forename: Xizhi
authors.initials: X
authors.lastname: Zhang
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.forename: Ming
authors.initials: M
authors.lastname: Ma
authors.forename: Shouhua
authors.initials: S
authors.lastname: Luo
authors.forename: Suyi
authors.initials: S
authors.lastname: Li
authors.forename: Ning
authors.initials: N
authors.lastname: Gu
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D015100:3,3'-Diaminobenzidine / Q000737:chemistry; D055162:Biocatalysis; D001943:Breast Neoplasms / Q000000981:diagnostic imaging / Q000378:metabolism; D045744:Cell Line, Tumor; D004396:Coloring Agents / Q000737:chemistry; D005260:Female; D006046:Gold; D006735:Horseradish Peroxidase; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods*; D053768:Metal Nanoparticles; D036103:Nanotechnology; D061848:Optical Imaging / Q000379:methods*; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000378:metabolism
chemical_list: D015100:3,3'-Diaminobenzidine; D004396:Coloring Agents; D006046:Gold; D006735:Horseradish Peroxidase; D018719:Erb-b2 Receptor Tyrosine Kinases; C508053:ERBB2 protein, human