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针对植物荧光遥感探测中信号易受干扰的问题, 提出了一种用于评估植物生长状况及环境监测的荧光寿命成像技术. 采用凹透镜对355 nm波长的激光扩束, 再照射植物激发叶绿素荧光, 由增强型电荷耦合器件接收荧光信号. 采用时间分辨测量法, 连续用相同激光脉冲照射植物以激发相同的荧光信号, 同时不断改变激光脉冲触发探测器启动的延时时间, 从而能够得到完整的离散荧光信号分布图像. 对植物特定位置点产生的离散荧光信号进行拟合, 再运用一种改进型的迭代解卷积法可反演高精度的荧光寿命; 进而反演图像各点的荧光寿命以生成植物的荧光寿命分布图. 该方法所绘制的荧光寿命图比荧光强度图能更准确地反映植物内部的叶绿素含量, 并对活体植物叶绿素荧光寿命的物理特性进行了初步研究, 证明叶绿素荧光寿命与植物生理状态存在一定关联; 并且叶绿素荧光寿命与活体植物所处环境存在着复杂的关系. 未来将与生物物理学家们合作, 继续探寻叶绿素荧光寿命与植物生存环境的关系.Plant fluorescence is a susceptible signal in plant fluorescence remote sensing detection. In order to solve this problem, a technique for plant chlorophyll fluorescence lifetime imaging is presented to evaluate living status for plant growth and environmental monitoring. A concave lens is used to expand laser beam at a wavelength of 355 nm, and the living plant is exposed in this laser light source to excite chlorophyll fluorescence. And the chlorophyll fluorescence signals are detected by an intensification charge coupled device. Time resolved measurement method is used in this article, so that every time the same fluorescence signals can be excited by the same laser pulse. Meanwhile, the delay time needed for triggering intensification charge coupled device should be changed consecutively, and the whole discrete fluorescence signal can be obtained. The discrete fluorescence signals from the particular location points of the plant are fitted. An improved method of forward iterative deconvolution is used to retrieve the corresponding fluorescence lifetime, and the high-precision fluorescence lifetime can be obtained. Furthermore, the fluorescence lifetime values at all the location points are retrieved to obtain the distribution map of chlorophyll fluorescence lifetime. This method can give the chlorophyll fluorescence image efficiently. The distribution map of fluorescence lifetime can more effectively reflect the plant chlorophyll concentration than the fluorescence intensity image does. The physical property of chlorophyll fluorescence lifetime from living plants has been studied preliminarily, indicating that the plant physiological status is related to its fluorescence lifetime to a certain extent; and the chlorophyll fluorescence lifetime and plant environment have a subtle and complex correlation. In the future, the relationship between chlorophyll fluorescence lifetime and plant environment will be expected to study with the cooperation of biophysicist.
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Keywords:
- fluorescence lifetime imaging /
- laser-induced fluorescence /
- fluorescence LiDAR /
- chlorophyll
[1] Janusauskaite D, Feiziene D 2002 Acta. Agr. Scand. B-S. P. 62 7
[2] Tol C, Verhoef W, Rosema A 2009 Agr. Forest Meteorol. 149 96
[3] Men Z W, Fang W H, Li Z W, Qu G N, Gao S Q, Lu G H, Yang J G, Sun C L 2010 Chin. Phys. B 19 8
[4] Fu C Y, Ng B K, Razul S G 2009 J. Biomed Opt. 14 064009
[5] Zhao M, Peng L L 2010 Opt. Lett. 35 2910
[6] Jordi R, Michael S, Santiago R 2012 Opt. Lett. 37 1229
[7] Hungerford G, Birch D J S 1996 Meas. Sci. Technol. 7 121
[8] Yuan S, Chin S L, Zeng H P 2015 Chin. Phys. B 24 1
[9] Roberts M S, Dancik Y, Prow T W, Thorling C A, Lin L L, Grice J E, Robertson T A, Konig K, Becker W 2011 Eur. J. Pharm Biopharm 77 469
[10] Gutierrez-Navarro O, Campos-Delgado D U, Arce-Santana E R, Maitland K C, Cheng S, Jabbour J, Malik B, Cuenca R, Jo J A 2014 Optics Express 22 12255
[11] Miao Z, Li S F, Zhang Q Y 2006 Acta Phys. Sin. 55 4321 (in Chinese) [苗壮, 李善锋, 张庆瑜 2006 55 4321]
[12] Alex S, Mary L 2014 Opt Lett 39 5362
[13] Oliveira F F, Ito A S, Bachmann L 2010 Appl. Optics 49 2244
[14] Wei H Y, Xu T, Wang F, Peng X S, Wei X, Liu S Y 2013 Acta Optica Sinica 33 0823001 (in Chinese) [魏惠月, 徐涛, 王峰, 彭晓世, 韦欣, 刘慎业 2013 光学学报 33 0823001]
[15] Garca M, Vzquez R, Isakina S, Lpez R 2012 Electrical Communications and Computers 27 317
[16] Kennifer R D, Peter M A T 2003 Practical applications of chlorophyll fluorescence in plant biology (Holland: Kluwer Academic Publishers) pp109-132
[17] Kotzagianni M, Couris S 2013 Chemical Physics Letters 561 36
[18] Xu M F, Shi Y B, Gao W H, Chen H Y 2014 Chinese Journal of Lasers 41 0108005 (in Chinese) [徐美芳, 石云波, 高文宏, 陈海洋 2014 中国激光 41 0108005]
[19] Mukamel E A, Babcock H, Zhuang X W 2012 Biophysical Journal 102 2391
[20] Wan W B, Hua D X, Le J, Liu M X, Cao N 2013 Acta Phys. Sin. 62 190601(in Chinese) [万文博, 华灯鑫, 乐静, 刘美霞, 曹宁 2013 62 190601]
[21] Lichtenthaler H K 2011 Applications of chlorophyll fluorescene (Holland: Kluwer Academic Publishers) pp182-196
[22] Kumar P J, Gopal R 2011 Spectroscopy-Biomedical Applications 26 129
[23] Fang W H, Li Z W, Li Z L, Qu G N, Ouyang S L, Men Z W 2012 Acta Phys. Sin. 61 153301(in Chinese) [房文汇, 里佐威, 李占龙, 曲冠男, 欧阳顺利, 门志伟 2012 61 153301]
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[1] Janusauskaite D, Feiziene D 2002 Acta. Agr. Scand. B-S. P. 62 7
[2] Tol C, Verhoef W, Rosema A 2009 Agr. Forest Meteorol. 149 96
[3] Men Z W, Fang W H, Li Z W, Qu G N, Gao S Q, Lu G H, Yang J G, Sun C L 2010 Chin. Phys. B 19 8
[4] Fu C Y, Ng B K, Razul S G 2009 J. Biomed Opt. 14 064009
[5] Zhao M, Peng L L 2010 Opt. Lett. 35 2910
[6] Jordi R, Michael S, Santiago R 2012 Opt. Lett. 37 1229
[7] Hungerford G, Birch D J S 1996 Meas. Sci. Technol. 7 121
[8] Yuan S, Chin S L, Zeng H P 2015 Chin. Phys. B 24 1
[9] Roberts M S, Dancik Y, Prow T W, Thorling C A, Lin L L, Grice J E, Robertson T A, Konig K, Becker W 2011 Eur. J. Pharm Biopharm 77 469
[10] Gutierrez-Navarro O, Campos-Delgado D U, Arce-Santana E R, Maitland K C, Cheng S, Jabbour J, Malik B, Cuenca R, Jo J A 2014 Optics Express 22 12255
[11] Miao Z, Li S F, Zhang Q Y 2006 Acta Phys. Sin. 55 4321 (in Chinese) [苗壮, 李善锋, 张庆瑜 2006 55 4321]
[12] Alex S, Mary L 2014 Opt Lett 39 5362
[13] Oliveira F F, Ito A S, Bachmann L 2010 Appl. Optics 49 2244
[14] Wei H Y, Xu T, Wang F, Peng X S, Wei X, Liu S Y 2013 Acta Optica Sinica 33 0823001 (in Chinese) [魏惠月, 徐涛, 王峰, 彭晓世, 韦欣, 刘慎业 2013 光学学报 33 0823001]
[15] Garca M, Vzquez R, Isakina S, Lpez R 2012 Electrical Communications and Computers 27 317
[16] Kennifer R D, Peter M A T 2003 Practical applications of chlorophyll fluorescence in plant biology (Holland: Kluwer Academic Publishers) pp109-132
[17] Kotzagianni M, Couris S 2013 Chemical Physics Letters 561 36
[18] Xu M F, Shi Y B, Gao W H, Chen H Y 2014 Chinese Journal of Lasers 41 0108005 (in Chinese) [徐美芳, 石云波, 高文宏, 陈海洋 2014 中国激光 41 0108005]
[19] Mukamel E A, Babcock H, Zhuang X W 2012 Biophysical Journal 102 2391
[20] Wan W B, Hua D X, Le J, Liu M X, Cao N 2013 Acta Phys. Sin. 62 190601(in Chinese) [万文博, 华灯鑫, 乐静, 刘美霞, 曹宁 2013 62 190601]
[21] Lichtenthaler H K 2011 Applications of chlorophyll fluorescene (Holland: Kluwer Academic Publishers) pp182-196
[22] Kumar P J, Gopal R 2011 Spectroscopy-Biomedical Applications 26 129
[23] Fang W H, Li Z W, Li Z L, Qu G N, Ouyang S L, Men Z W 2012 Acta Phys. Sin. 61 153301(in Chinese) [房文汇, 里佐威, 李占龙, 曲冠男, 欧阳顺利, 门志伟 2012 61 153301]
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