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

Affiliations
  • 1Department of Surgery, Catholic University of Daegu School of Medicine, Daegu, Korea. shwpark@cu.ac.kr
  • 2Department of Anatomy, Catholic University of Daegu School of Medicine, Daegu, Korea.
  • 3Department of Radiology and Biomedical Engineering, Catholic University of Daegu School of Medicine, Daegu, Korea.
  • 4Department of Pathology, Catholic University of Daegu School of Medicine, Daegu, Korea.
  • 5Pohang Accelerator Laboratory, Pohang University of Science and Technology, Pohang, Korea.

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.


Reference

1. Housa D, Housová J, Vernerová Z, Haluzik M. Adipocytokines and cancer. Physiol Res. 2006. 55:233–244.
Article
2. Jatoi I, Miller AB. Why is breast cancer mortality declining? Lancet Oncol. 2003. 4:251–254.
3. Margaritondo G, Meuli R. Synchrotron radiation in radiology: novel X-ray sources. Eur Radiol. 2003. 13:2633–2641.
Article
4. Lewis RA, Rogers KD, Hall CJ, Hufton AP, Evans S, Menk RH, et al. Antonuk LE, Yaffe MJ, editors. Diffraction enhanced imaging: improved contrast, lower dose Xray imaging. Medical Imaging 2002: Physics of Medical Imaging, San Diego, USA. 2002. Bellingham: Society of Photo-optical Instrumentation Engineers;286–297.
5. Lewis RA. Meidcal phase contrast X-ray imaging: current status and future prospects. Phys Med Biol. 2004. 49:3573–3583.
6. Suortti P, Thomlinson W. Medical applications of synchrotron radiation. Phys Med Biol. 2003. 48:R1–R35.
Article
7. Burattini E, Cossu E, Maggio C, Gambaccini M, Indovina PL, Marziani M, et al. Mammography with synchrotron radiation. Radiology. 1995. 195:239–244.
Article
8. Arfelli F, Bonvicini V, Bravin A, Cantatore G, Castelli E, Palma LD, et al. Mammography with synchrotron radiation: phase-detection techniques. Radiology. 2000. 215:286–293.
Article
9. Ingal VN, Beliaevskaya EA, Brianskaya AP, Merkurieva RD. Phase mammography: a new technique for breast investigation. Phys Med Biol. 1998. 43:2555–2567.
10. Liu C, Yan X, Zhang X, Yang W, Peng W, Shi D, et al. Evaluation of X-ray diffraction enhanced imaging in the diagnosis of breast cancer. Phys Med Biol. 2007. 52:419–427.
Article
11. Fiedler S, Bravin A, Keyriläinen J, Fernández M, Suortti P, Thomlinson W, et al. Imaging lobular breast carcinoma: comparison of synchrotron radiation DEI-CT technique with clinical CT, mammography and histology. Phys Med Biol. 2004. 49:175–188.
Article
12. Jeong YJ, Bong JG, Kim HT, Kim JK, Jheon SH, Youn HS, et al. Synchrotron radiation imaging of female breast tissues using phase contrast technique. J Breast Cancer. 2008. 11:40–44.
Article
13. Park SH, Kim HT, Kim JK, Jheon SH, Youn HS. Hard X-ray microscopic imaging of human breast tissues. 2006. In : 9th International Conference on Synchrotron Radiation Instrumentation; Daegu: American Institute of Physics.
14. Stuart JS, Anthony JG. Harris JR, Lippman ME, Morrow M, Osborne CK, editors. Pathology of invasive breast cancer. Diseases of the Breast. 2004. 3rd ed. Philadelphia: Lippincott Williams & Wilkins;542–584.
15. Azzopardi JG. Azzopardi JG, Ahmed A, Millis RR, editors. Epitheliosis and in situ carcinoma. Problems in Breast Pathology. 1979. Philadelphia: Saunders;128.
16. Rosen PP. Rosen PP, editor. Paget's disease of the nipple. Rosen' Breast Pathology. 1997. Philadelphia: Lippincott-Raven;493–506.
17. Youn HS, Jung SW. Hard X-ray microscopy with Zernike phase contrast. J Microsc. 2006. 223(Pt 1):53–56.
Article
18. Weon BM, Je JH, Hwu Y, Margaritondo G. Phase contrast X-ray imaging. Int J Nanotechnol. 2006. 3:280–297.
Article
19. Socha JJ, Westneat MW, Harrison JF, Waters JS, Lee WK. Real-time phase-contrast X-ray imaging: a new technique for the study of animal form and function. BMC Biol. 2007. 5:6.
Article
20. Kagoshima Y, Yokoyama Y, Niimi T, Koyama T, Tsusaka Y, Matsui J, et al. Hard X-ray phase-contrast microscope for observing transparent specimens. 2002. In : 7th International Conference on X-Ray Microscopy; Grenoble: European Synchrotron Radiation Facility.
21. Andrews JC, Almeida E, van der Meulen MC, Alwood JS, Lee C, Liu Y, et al. Nanoscale X-ray microscopic imaging of mammalian mineralized tissue. Microsc Microanal. 2010. 16:327–336.
Article
22. Chen J, Yang Y, Zhang X, Antrews JC, Pianetta P, Guan Y, et al. 3D nanoscale imaging of the yeast, Schizosaccharomyces pombe, by full-field transmission X-ray microscopy at 5.4 keV. Anal Bioanal Chem. 2010. 397:2117–2121.
Article
23. Youn HS, Baik SY, Chang CH. Hard X-ray microscopy with a 130 nm spatial resolution. Rev Sci Instrum. 2005. 76:023702.
24. Schiffhauer LM, Boger JN, Bonfiglio TA, Zavislan JM, Zuley M, Fox CA. Confocal microscopy of unfixed breast needle core biopsies: a comparison to fixed and stained sections. BMC Cancer. 2009. 9:265.
Article
25. Huang D, Swanson EA, Lin CP, Schuman JS, Stinson WG, Chang W, et al. Optical coherence tomography. Science. 1991. 254:1178–1181.
Article
26. Fercher AF, Drexler W, Hitzenberger CK, Lasser T. Optical coherence tomography: principles and applications. Rep Prog Phys. 2003. 66:239–303.
27. Pani S, Longo R, Derossi D, Montanari F, Olivo A, Arfelli F, et al. Breast tomography with synchrotron radiation: preliminary results. Phys Med Biol. 2004. 49:1739–1754.
Article
28. Lindfors KK, Boone JM, Nelson TR, Yang K, Kwan AL, Miller DF. Dedicated breast CT: initial clinical experience. Radiology. 2008. 246:725–733.
Article
29. Arfelli F, Assante M, Bonvicini V, Bravin A, Cantatore G, Castelli E, et al. Low-dose phase contrast X-ray medical imaging. Phys Med Biol. 1998. 43:2845–2852.
Article
30. Boone JM, Nelson TR, Lindfors KK, Seibert JA. Dedicated breast CT: radiation dose and image quality evaluation. Radiology. 2001. 221:657–667.
Article
Full Text Links
  • JBC
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr