光片荧光显微镜长时间的稳定成像
|
摘要
利用4f(f为焦距)系统和电子耦合器件相机跟踪样品的位置,通过纳米平移台对机械漂移进行了补偿,使样品始终处于光片的束腰位置,从而获得了最优图像。所提方法可以对几纳米的机械偏移进行补偿。使用自主搭建的光片荧光显微镜对直径为几微米的荧光微球进行了长时间成像,验证了所提方法的可行性。
By using a 4f(f is the focal length) optical setup and a charge coupled device camera, the sample position is tracked. The mechanical drift is compensated by the nano-translation table which ensures the sample always at the beam waist of the light sheet and thus an optimal image is obtained. With the proposed method, the mechanical drift of several nanometers can be compensated. By using the home-made light sheet fluorescence microscopy, the long-time imaging of fluorescent microspheres with diameters of several micrometers is acquired, which confirms the feasibility of the proposed method.
By using a 4f(f is the focal length) optical setup and a charge coupled device camera, the sample position is tracked. The mechanical drift is compensated by the nano-translation table which ensures the sample always at the beam waist of the light sheet and thus an optimal image is obtained. With the proposed method, the mechanical drift of several nanometers can be compensated. By using the home-made light sheet fluorescence microscopy, the long-time imaging of fluorescent microspheres with diameters of several micrometers is acquired, which confirms the feasibility of the proposed method.
引文
[1] Tokunaga M,Imamoto N,Sakata-Sogawa K.Highly inclined thin illumination enables clear single-molecule imaging in cells[J].Nature Methods,2008,5(2):159-161.
[2] Tomer R,Khairy K,Keller P J.Light sheet microscopy in cell biology[M]//Tomer R,Khairy K,Keller P J.eds.Methods in molecular biology.Totowa,New Jersey:Humana Press,2012:123-137.
[3] Zickus V,Taylor J M.3D+time blood flow mapping using SPIM-microPIV in the developing zebrafish heart[J].Biomedical Optics Express,2018,9(5):2418-2435.
[4] Schmid B,Shah G,Scherf N,et al.High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics[J].Nature Communications,2013,4:2207.
[5] Haslehurst P,Yang Z Y,Dholakia K,et al.Fast volume-scanning light sheet microscopy reveals transient neuronal events[J].Biomedical Optics Express,2018,9(5):2154-2167.
[6] Keller P J,Ahrens M B,Freeman J.Light-sheet imaging for systems neuroscience[J].Nature Methods,2015,12(1):27-29.
[7] Wu Y C,Kumar A,Smith C,et al.Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy[J].Nature Communications,2017,8:1452.
[8] Hedde P N,Gratton E.Active focus stabilization for upright selective plane illumination microscopy[J].Optics Express,2015,23(11):14707-14714.
[9] An K,Wang J,Liang D,et al.Improving lateral resolution of light sheet fluorescence microscopy with SOFI method[J].Chinese Journal of Lasers,2017,44(6):0607002.安坤,王晶,梁东,等.利用SOFI方法提高光片荧光显微镜横向分辨率[J].中国激光,2017,44(6):0607002.
[10] Gao L,Shao L,Chen B C,et al.3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy[J].Nature Protocols,2014,9(5):1083-1101.
[11] Olarte O E,Andilla J,Gualda E J,et al.Light-sheet microscopy:a tutorial[J].Advances in Optics and Photonics,2018,10(1):111-179.
[12] Yang Y L,Zong W J,Wu R L,et al.Light-sheet fluorescence microscopy[J].Acta Optica Sinica,2017,37(3):0318007.杨豫龙,宗伟建,吴润龙,等.光片荧光显微成像[J].光学学报,2017,37(3):0318007.
[13] Planchon T A,Gao L,Milkie D E,et al.Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination[J].Nature Methods,2011,8(5):417-423.
[14] Power R M,Huisken J.A guide to light-sheet fluorescence microscopy for multiscale imaging[J].NatureMethods,2017,14(4):360-373.
[15] Reynaud E G,Peychl J,Huisken J,et al.Guide to light-sheet microscopy for adventurous biologists[J].Nature Methods,2015,12(1):30-34.
[16] Fahrbach F O,Voigt F F,Schmid B,et al.Rapid 3D light-sheet microscopy with a tunable lens[J].Optics Express,2013,21(18):21010-21026.
[17] Yoshida A,Kaburagi Y,Hishiyama Y.Full width at half maximum intensity of G band in first order Raman spectrum of carbon material as a parameter for graphitization-a study with pyrolytic carbons[J].Tanso,2006,2006(221):2-7.
[2] Tomer R,Khairy K,Keller P J.Light sheet microscopy in cell biology[M]//Tomer R,Khairy K,Keller P J.eds.Methods in molecular biology.Totowa,New Jersey:Humana Press,2012:123-137.
[3] Zickus V,Taylor J M.3D+time blood flow mapping using SPIM-microPIV in the developing zebrafish heart[J].Biomedical Optics Express,2018,9(5):2418-2435.
[4] Schmid B,Shah G,Scherf N,et al.High-speed panoramic light-sheet microscopy reveals global endodermal cell dynamics[J].Nature Communications,2013,4:2207.
[5] Haslehurst P,Yang Z Y,Dholakia K,et al.Fast volume-scanning light sheet microscopy reveals transient neuronal events[J].Biomedical Optics Express,2018,9(5):2154-2167.
[6] Keller P J,Ahrens M B,Freeman J.Light-sheet imaging for systems neuroscience[J].Nature Methods,2015,12(1):27-29.
[7] Wu Y C,Kumar A,Smith C,et al.Reflective imaging improves spatiotemporal resolution and collection efficiency in light sheet microscopy[J].Nature Communications,2017,8:1452.
[8] Hedde P N,Gratton E.Active focus stabilization for upright selective plane illumination microscopy[J].Optics Express,2015,23(11):14707-14714.
[9] An K,Wang J,Liang D,et al.Improving lateral resolution of light sheet fluorescence microscopy with SOFI method[J].Chinese Journal of Lasers,2017,44(6):0607002.安坤,王晶,梁东,等.利用SOFI方法提高光片荧光显微镜横向分辨率[J].中国激光,2017,44(6):0607002.
[10] Gao L,Shao L,Chen B C,et al.3D live fluorescence imaging of cellular dynamics using Bessel beam plane illumination microscopy[J].Nature Protocols,2014,9(5):1083-1101.
[11] Olarte O E,Andilla J,Gualda E J,et al.Light-sheet microscopy:a tutorial[J].Advances in Optics and Photonics,2018,10(1):111-179.
[12] Yang Y L,Zong W J,Wu R L,et al.Light-sheet fluorescence microscopy[J].Acta Optica Sinica,2017,37(3):0318007.杨豫龙,宗伟建,吴润龙,等.光片荧光显微成像[J].光学学报,2017,37(3):0318007.
[13] Planchon T A,Gao L,Milkie D E,et al.Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination[J].Nature Methods,2011,8(5):417-423.
[14] Power R M,Huisken J.A guide to light-sheet fluorescence microscopy for multiscale imaging[J].NatureMethods,2017,14(4):360-373.
[15] Reynaud E G,Peychl J,Huisken J,et al.Guide to light-sheet microscopy for adventurous biologists[J].Nature Methods,2015,12(1):30-34.
[16] Fahrbach F O,Voigt F F,Schmid B,et al.Rapid 3D light-sheet microscopy with a tunable lens[J].Optics Express,2013,21(18):21010-21026.
[17] Yoshida A,Kaburagi Y,Hishiyama Y.Full width at half maximum intensity of G band in first order Raman spectrum of carbon material as a parameter for graphitization-a study with pyrolytic carbons[J].Tanso,2006,2006(221):2-7.