J Breast Cancer.  2010 Dec;13(4):349-356. 10.4048/jbc.2010.13.4.349.

Evaluation of Phase-Contrast Microscopic Imaging with Synchrotron Radiation in the Diagnosis of Breast Cancer and Differentiation of Various Breast Diseases: Preliminary Results

• Jeong YJ
• Bong JG
• Kim HT
• Kim JK
• Oh HK
• Huang JY
• Park SH

• KMID: 2286522
• DOI: http://doi.org/10.4048/jbc.2010.13.4.349

Abstract

PURPOSE
A significant improvement of imaging using synchrotron radiation (SR) is obtained by introducing phase-contrast technique. This technique provides greatly enhanced contrast and good soft tissue discrimination with high spatial resolution. The aim of this study was to observe microstructures of pathologic breast specimens including invasive breast cancer using phase-contrast technique with SR and to evaluate the feasibility of phase-contrast imaging in clinical application.
METHODS
Phase-contrast microscopic image of normal breast tissue and the images of various breast diseases such as fibrocystic change, ductal carcinoma in situ, invasive ductal carcinoma, Paget's disease were obtained using hard X-ray microscopy with an 11.1 keV monochromatic beam from SR source and CsI (TI) scintillation crystal. Zernike phase-shifter was adapted for phase-contrast hard X-ray microscopy. The visual image was magnified 20 times by microscopic objective lens and captured using a full frame charge-coupled device camera. Obtained images were compared with corresponding histopathologic findings in the optical microscopy.
RESULTS
The SR images of various breast diseases were obtained with a good contrast and high visibility by phase-contrast technique. It was possible to observe the microstructures with high spatial resolution down to the micron region. The characteristic features of each disease were consistent with the histopathologic findings of corresponding sample and the images of breast cancer and the other diseases were distinct from each other.
CONCLUSION
Using phase-contrast technique, SR images of various breast diseases including breast cancer were obtained. These images were comparable with standard histopathologic findings and showed different features for each disease. The results suggest that phase-contrast microscopic imaging with SR has potential as a diagnostic tool and also its clinical application is feasible, especially in breast imaging.

Keyword

Breast; Phase-contrast microscopy; Synchrotron radiation

MeSH Terms

Breast
Breast Diseases
Breast Neoplasms
Carcinoma, Ductal
Carcinoma, Intraductal, Noninfiltrating
Discrimination (Psychology)
Microscopy
Microscopy, Phase-Contrast
Synchrotrons

Figure

• 

  Figure 1 Schematic drawing of the experimental set-up. Monochromatic beam set at 11.1 keV (=50Å) irradiated the sample positioned 25 m away from the source. A visual signal on the surface of CsI (TI) scintillation crystal placed at a distance of 5 cm from specimen was magnified using a 20× microscopic objective lens and captured using CCD camera.

• 

  Figure 2 (A) Comparison of normal terminal duct-lobular unit (TDLU) structures of breast between synchrotron radiation (SR) image and (B) histologic section (H&E stain, ×40). SR image shows well defined TDLU surrounded by trabeculae of collagen bundles (*) and mature fat cells (arrow heads). Fine structures of small acini and interacinar spaces (arrow) of the TDLU are visible in SR image.

• 

  Figure 3 (A) Phase-contrast synchrotron radiation (SR) image and (B) optical microscopic image (H&E stain, ×40) of fibrocystic change. It shows cystic change (arrow) and fatty tissue (arrow head). 10 µm-thick formalin-fixed tissue was used and measured 1.3×0.9 mm in size.

• 

  Figure 4 (A) Comparison of synchrotron radiation (SR) image and (B) histologic section (H&E stain, ×40) of low grade ductal carcinoma in situ lesion of cribriform pattern. The SR image well correlated to the histologic section with prominent periductal basement membrane (arrows) and stippled fine microcalcifications (black dots in left tumor nest, circle). Round fenestrae (*) of the tumor cell nests are highlighted in the SR image.

• 

  Figure 5 (A) Comparison of synchrotron radiation (SR) image and (B) histologic section of ductal carcinoma in situ lesion of solid and cribriform pattern (H&E stain, ×40). The SR image well correlates to the histologic section with prominent periductal basement membrane (arrows) and stippled fine microcalcifications (black dots in tumor nest, arrow head). Peritumoral inflammatory cell infiltration are highlighted in the SR image (circle).

• 

  Figure 6 (A) Phase-contrast synchrotron radiation (SR) image and (B) optical microscopic image (H&E stain, ×40) of breast cancer. It shows irregular tumor cell infiltration into adjacent fat tissue (arrows) and parenchyme with severe stromal fibrosis (arrow head). (C) Cords and individual infiltrating tumor cells in the desmoplastic stroma are seen in a high-power field view (H&E stain, ×400). The specimen was formalin-fixed breast cancer tissue, 10 µm thick and 0.9×0.8 mm in size.

• 

  Figure 7 (A) Phase-contrast synchrotron radiation (SR) image and (B) optical microscopic image (H&E stain, ×40) of Paget's disease. SR image shows lacuna shaped intraepidermal Paget cells (arrows) of nipple epidermis. In SR image, the Paget cells show large electron dense nuclei and electron lucent abundant cytoplasm with distinct cell outlines, and most squamous cells are indistinct in cell border.

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