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提出了一种基于偏振滤波图像增强和动态散斑照明的新型宽场荧光层析显微镜. 该显微镜采用了一种新型的偏振滤波图像增强技术, 基于激发光与荧光偏振态的差异, 利用偏振器件滤除激发光; 并利用动态散斑照明实现宽场层析. 该荧光层析显微镜具有结构简单、低成本、响应速度快、容易操作等特点. 实验研究结果表明, 本文提出的滤波方案能够显著地提高图像质量, 利用动态散斑照明实现宽场层析具有较高的纵向分辨能力. 研究丰富了在荧光显微镜中, 从强激发光中提取弱荧光信号的技术手段, 为今后发展具有快速响应, 波长可调谐的多光谱荧光层析等高端的显微镜具有重要参考意义.We propose a novel wide-field fluorescence sectioning microscope based on polarization filtering image enhancement and dynamic speckle illumination in this article. A new type of polarization filtering image enhancement technology, based on the differences in polarization characteristic between excitation light and fluorescence, is used to filter out the excitation light and enhance image quality. The dynamic speckle illumination is employed to achieve the wide field fluorescence sectioning which has many advantages, such as simple configuration, low cost, rapid response and easy operation. The experimental results indicate that our filtering method can be utilized to improve the image quality significantly, and the dynamic using speckle illumination can be employed to achieve optical sectioning with a wide field and a high vertical resolution capability. Our study is not only to enrich the technological method to extract the weak fluorescence signal from the strong excitation light, but also to provide a significant reference for developing a light wavelength tunable multi-spectral fluorescence microscope with rapid response.
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Keywords:
- laser speckle /
- polarization filtering /
- fluorescence microscope /
- optical sectioning
[1] Wilson T, Shepard C J 1984 Theory and Practice of Scanning Optical Microscopy (New York: Academic Press)
[2] Denk W, Strickler J H, Webb W W 1990 Science 248 73
[3] Somekh M G, See C W, Goh J 2000 Opt. Commun. 2 75
[4] Braeckmans K, Peeters L, Sanders N N, De Smedt 2003 Bio. Phys. J. 85 2240
[5] Ventalon C, Mertz J 2005 Opt. Lett. 30 3350
[6] Ventalon C, Mertz J 2006 Opt. Express 14 7198
[7] Lin H M, Shao Y H, Qu J L, Yin J, Chen S P, Niu H B 2008 Acta Phys. Sin. 57 7641 (in Chinese) [林浩铭, 邵永红, 屈军乐, 尹君, 陈思平, 牛憨笨 2008 物理学报 57 7641]
[8] Shao Y H, Li H, Wen Q, Wang Y, Qu J L, Niu H B 2010 Chin. Opt. Lett. 10 944
[9] Lim D, Ford T N, Chu K K, Mertz J 2011 J. Biomed. Opt. 16 016014
[10] Mazzaferri J, Kunik D, Belisle J M, Singh K, Lefrancois S, Costantino S 2011 Opt. Express 19 14508
[11] Wen Q, Qu J L, Shao Y H, Wang K G, Niu H B China Patent 201010616211. X [2010-12-30]
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[1] Wilson T, Shepard C J 1984 Theory and Practice of Scanning Optical Microscopy (New York: Academic Press)
[2] Denk W, Strickler J H, Webb W W 1990 Science 248 73
[3] Somekh M G, See C W, Goh J 2000 Opt. Commun. 2 75
[4] Braeckmans K, Peeters L, Sanders N N, De Smedt 2003 Bio. Phys. J. 85 2240
[5] Ventalon C, Mertz J 2005 Opt. Lett. 30 3350
[6] Ventalon C, Mertz J 2006 Opt. Express 14 7198
[7] Lin H M, Shao Y H, Qu J L, Yin J, Chen S P, Niu H B 2008 Acta Phys. Sin. 57 7641 (in Chinese) [林浩铭, 邵永红, 屈军乐, 尹君, 陈思平, 牛憨笨 2008 物理学报 57 7641]
[8] Shao Y H, Li H, Wen Q, Wang Y, Qu J L, Niu H B 2010 Chin. Opt. Lett. 10 944
[9] Lim D, Ford T N, Chu K K, Mertz J 2011 J. Biomed. Opt. 16 016014
[10] Mazzaferri J, Kunik D, Belisle J M, Singh K, Lefrancois S, Costantino S 2011 Opt. Express 19 14508
[11] Wen Q, Qu J L, Shao Y H, Wang K G, Niu H B China Patent 201010616211. X [2010-12-30]
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