Quantitative diagnostic imaging of cancer tissues by using phosphor-integrated dots with ultra-high brightness

Sci Rep. 2017 Aug 8;7(1):7509. doi: 10.1038/s41598-017-06534-z.

Abstract

The quantitative sensitivity and dynamic range of conventional immunohistochemistry (IHC) with 3,3'-diaminobenzidine (IHC-DAB) used in pathological diagnosis in hospitals are poor, because enzyme activity can affect the IHC-DAB chromogenic reaction. Although fluorescent IHC can effectively increase the quantitative sensitivity of conventional IHC, tissue autofluorescence interferes with the sensitivity. Here, we created new fluorescent nanoparticles called phosphor-integrated dots (PIDs). PIDs have 100-fold greater brightness and a more than 300-fold greater dynamic range than those of commercially available fluorescent nanoparticles, quantum dots, whose fluorescence intensity is comparable to tissue autofluorescence. Additionally, a newly developed image-processing method enabled the calculation of the PID particle number in the obtained image. To quantify the sensitivity of IHC using PIDs (IHC-PIDs), the IHC-PIDs method was compared with fluorescence-activated cell sorting (FACS), a method well suited for evaluating total protein amount, and the two values exhibited strong correlation (R = 0.94). We next applied IHC-PIDs to categorize the response to molecular target-based drug therapy in breast cancer patients. The results suggested that the PID particle number estimated by IHC-PIDs of breast cancer tissues obtained from biopsy before chemotherapy can provide a score for predicting the therapeutic effect of the human epidermal growth factor receptor 2-targeted drug trastuzumab.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3,3'-Diaminobenzidine / chemistry
  • Antibodies / chemistry
  • Antineoplastic Agents, Immunological / therapeutic use
  • Biopsy
  • Biotin / chemistry
  • Breast Neoplasms / diagnostic imaging*
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / genetics
  • Breast Neoplasms / pathology
  • Diagnostic Imaging / instrumentation
  • Diagnostic Imaging / methods*
  • Female
  • Fluorescence
  • Fluorescent Dyes / chemistry*
  • Gene Expression
  • Humans
  • Imides / chemistry
  • Immunohistochemistry / methods
  • Middle Aged
  • Nanoparticles / chemistry*
  • Neoplasm Proteins / antagonists & inhibitors
  • Neoplasm Proteins / genetics
  • Neoplasm Proteins / metabolism
  • Particle Size
  • Perylene / analogs & derivatives
  • Perylene / chemistry
  • Receptor, ErbB-2 / antagonists & inhibitors
  • Receptor, ErbB-2 / genetics
  • Receptor, ErbB-2 / metabolism
  • Rhodamines / chemistry*
  • Streptavidin / chemistry
  • Trastuzumab / therapeutic use

Substances

  • Antibodies
  • Antineoplastic Agents, Immunological
  • Fluorescent Dyes
  • Imides
  • Neoplasm Proteins
  • Rhodamines
  • perylene bisimide
  • 3,3'-Diaminobenzidine
  • Perylene
  • tetramethylrhodamine
  • Biotin
  • Streptavidin
  • ERBB2 protein, human
  • Receptor, ErbB-2
  • Trastuzumab