Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles

Journal Article · Fri Sep 28 04:00:00 EDT 2012 · Biochemical and Biophysical Research Communications

DOI:https://doi.org/10.1016/J.BBRC.2012.08.105· OSTI ID:22210265

Gonda, Kohsuke ^[1]; Miyashita, Minoru ^[2]; Watanabe, Mika; Takahashi, Yayoi ^[3]; Goda, Hideki; Okada, Hisatake; Nakano, Yasushi ^[4]; Tada, Hiroshi ^[2]; Amari, Masakazu ^[1]; Ohuchi, Noriaki ^[1]

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1. Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8575 (Japan)
2. Department of Surgical Oncology, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8574 (Japan)
3. Department of Pathology, Tohoku University Hospital, Seiryo-machi, Aoba-ku, Sendai 980-8574 (Japan)
4. Optical and Biological R and D Center, Konica Minolta Technology Center, Inc., No. 1 Sakuramachi, Hino-shi, Tokyo 191-8511 (Japan)

Highlights: Black-Right-Pointing-Pointer Organic fluorescent material-assembled nanoparticles for IHC were prepared. Black-Right-Pointing-Pointer New nanoparticle fluorescent intensity was 10.2-fold greater than Qdot655. Black-Right-Pointing-Pointer Nanoparticle staining analyzed a wide range of ER expression levels in tissue. Black-Right-Pointing-Pointer Nanoparticle staining enhanced the quantitative sensitivity for ER diagnosis. -- Abstract: The detection of estrogen receptors (ERs) by immunohistochemistry (IHC) using 3,3 Prime -diaminobenzidine (DAB) is slightly weak as a prognostic marker, but it is essential to the application of endocrine therapy, such as antiestrogen tamoxifen-based therapy. IHC using DAB is a poor quantitative method because horseradish peroxidase (HRP) activity depends on reaction time, temperature and substrate concentration. However, IHC using fluorescent material provides an effective method to quantitatively use IHC because the signal intensity is proportional to the intensity of the photon excitation energy. However, the high level of autofluorescence has impeded the development of quantitative IHC using fluorescence. We developed organic fluorescent material (tetramethylrhodamine)-assembled nanoparticles for IHC. Tissue autofluorescence is comparable to the fluorescence intensity of quantum dots, which are the most representative fluorescent nanoparticles. The fluorescent intensity of our novel nanoparticles was 10.2-fold greater than quantum dots, and they did not bind non-specifically to breast cancer tissues due to the polyethylene glycol chain that coated their surfaces. Therefore, the fluorescent intensity of our nanoparticles significantly exceeded autofluorescence, which produced a significantly higher signal-to-noise ratio on IHC-imaged cancer tissues than previous methods. Moreover, immunostaining data from our nanoparticle fluorescent IHC and IHC with DAB were compared in the same region of adjacent tissues sections to quantitatively examine the two methods. The results demonstrated that our nanoparticle staining analyzed a wide range of ER expression levels with higher accuracy and quantitative sensitivity than DAB staining. This enhancement in the diagnostic accuracy and sensitivity for ERs using our immunostaining method will improve the prediction of responses to therapies that target ERs and progesterone receptors that are induced by a downstream ER signal.

• OSTI ID: 22210265
• Journal Information: Biochemical and Biophysical Research Communications, Journal Name: Biochemical and Biophysical Research Communications Journal Issue: 3 Vol. 426; ISSN BBRCA9; ISSN 0006-291X
• Country of Publication: United States
• Language: English

Related Subjects

60 APPLIED LIFE SCIENCES
ACCURACY
ANIMAL TISSUES
ERBIUM SULFIDES
ESTROGENS
FLUORESCENCE
MAMMARY GLANDS
NEOPLASMS
PEROXIDASES
POLYETHYLENE GLYCOLS
PROGESTERONE
RECEPTORS
SENSITIVITY
SIGNAL-TO-NOISE RATIO
TAMOXIFEN