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大视场光学显微成像技术

王义强 林方睿 胡睿 刘丽炜 屈军乐

王义强, 林方睿, 胡睿, 刘丽炜, 屈军乐. 大视场光学显微成像技术[J]. 中国光学(中英文). doi: 10.37188/CO.2022-0098
引用本文: 王义强, 林方睿, 胡睿, 刘丽炜, 屈军乐. 大视场光学显微成像技术[J]. 中国光学(中英文). doi: 10.37188/CO.2022-0098
WANG Yi-qiang, LIN Fang-rui, HU Rui, LIU Li-wei, QU Jun-le. Large field-of-view optical microscopic imaging technology[J]. Chinese Optics. doi: 10.37188/CO.2022-0098
Citation: WANG Yi-qiang, LIN Fang-rui, HU Rui, LIU Li-wei, QU Jun-le. Large field-of-view optical microscopic imaging technology[J]. Chinese Optics. doi: 10.37188/CO.2022-0098

大视场光学显微成像技术

doi: 10.37188/CO.2022-0098
详细信息
    作者简介:

    王义强(1996—),男,安徽合肥人,深圳大学硕士生,主要从事荧光显微成像系统的研究。E-mail:1656335032@qq.com

    林方睿(1993—),男,福建武夷山人,深圳大学博士生,主要从事荧光显微成像系统的研究。E-mail:lfr1993@163.com

    胡 睿(1983—),男,九三学社社员,现任深圳大学物理与光电工程学院副教授。2004年获浙江大学信息工程学士学位,2010年获浙江大学光学工程博士学位。2012~2015年于新加坡南洋理工大学从事博士后研究。主要从事生物医学光子学相关领域的研究工作,近年来专注于纳米颗粒与细胞的相互作用,通过光学成像及光谱学分析等方法研究二者相互作用过程的细节问题。邮箱:rhu@szu.edu.cn

    刘丽炜(1980—)女,吉林,博士,教授,2003年获长春理工大学学士,2009年获长春理工大学硕士,2013年获长春理工大学博士。研究方向:生物医学光子学,邮箱:liulw@szu.edu.cn

  • 中图分类号: O435.2;O438.2

Large field-of-view optical microscopic imaging technology

More Information
  • 摘要:

    光学显微成像技术具有实时、高分辨率和非侵入性等特点,其成像尺度可跨越细胞、组织乃至生命体,极大地拓展了人们对生命本质的认识边界。然而,受限于光学显微成像系统有限的空间带宽积(Space-bandwidth Product,SBP),常规的光学显微镜难以同时兼顾视场大小和分辨率,使得显微成像在大视场生物成像应用中受到较大的限制,例如对脑神经网络以突触为单位的神经回路成像。近年来,大视场光学显微成像技术得到不断的发展,其SBP相较于传统的光学显微镜有了十倍甚至百倍的提升,在保持高分辨率的基础上拓展了成像视场,满足了生物医学领域重大问题的研究需求。本文介绍了近年来几种典型的大视场光学显微成像技术及其生物医学应用,并对其未来发展做了展望。

     

  • 图 1  大视场物镜双光子脑成像[12]。(a)成像系统光路图和物镜实物图(左上);(b) 活体鼠脑神经细胞的双光子成像,深度为150 μm;(c) (b)中虚线框内的细节放大图。

    Figure 1.  Large FOV objective two-photon brain imaging[12]. (a) Objective and imaging system; (b) Two-photon imaging of neuronal cells activity of mice brain in vivo, the depth of imaging is 150 μm; (c) Magnification of some details in white box of (b).

    图 2  基于散斑光片照明的大视场成像系统[25]。(a) 系统光路示意图,激光经ground glass disk形成的散斑图案投影在体积为4.4×3×3 mm3的样品内,形成厚度约为3 μm的散斑光片照明;(b) 用光片照明模式实现斑马鱼全身成像;(c)用共聚焦模式实现的斑马鱼全身成像。

    Figure 2.  Speckle light sheet illumination-based large FOV imaging system[25]. (a) System setting, the speckle pattern formed by the laser through the ground glass disk is projected into the sample with a volume of 4.4×3×3 mm3, forming a speckle light sheet with a thickness of about 3 μm for illumination; (b) Zebrafish whole body imaging with light sheet illumination; (c) Zebrafish whole body imaging with confocal microscopy.

    图 3  微透镜的排列与扫描方向示意[37]

    Figure 3.  Schematic diagram of microlenses arrangement and scanning direction.

    图 4  阵列显微系统[39]。 (a) 阵列显微系统光路设置;(b) 小鼠肾脏切片的高通量成像;(c) (b)中红色框内的放大图。

    Figure 4.  Array microscopy[39]. (a) The optical setting of array microscopy; (b) High-throughput imaging of mouse kidney slices; (c) Magnification of the red rectangle in (b).

    图 5  无透镜分辨率增强成像系统[21]。(a) 系统结构和其使用的无序表面示意图;(b) 系统对小鼠肾脏切片的高通量成像,并选取三个子区域b1, b2, b3(图中红色框内)的成像结果与使用20 x, 0.75 NA物镜的荧光显微镜成像结果对比,两者的相似度约为0.75。

    Figure 5.  Resolution enhanced lensless imaging system[21]. (a) System design and the disordered surface; (b) High-throughput imaging of mouse kidney slices, and compared the imaging results of three sub-regions b1, b2, b3 (red boxes) with the imaging results of the fluorescence microscope using an objective with 20 x and 0.75 NA. The similarity is about 0.75.

    表  1  四类典型的大视场光学显微成像技术参数对比

    Table  1.   Comparison of four representative large FOV optical microscopy imaging techniques

    技术类别成像方式分辨率帧率(fps)视场优缺点适用场景
    大视场物镜成像宽场[27]0.7 μm9220 mm2可兼容多种成像方式;像质分布不均匀活体、细胞、切片观察
    双光子[17,27]0.6 μm5×10−320 mm2
    光片[29]0.7 μm0.1520 mm2
    曲面探测成像串行成像[37]1.5 μm0.71256 mm2整体像质更好;成像方式多局限于宽场活体、细胞、切片观察
    并行成像[19]1.2 μm30113 mm2
    阵列显微宽场+扫描[41]1.7 μm660 mm2简单;焦深浅切片观测
    无透镜显微多波长复用[53]0.69 μm0.529 mm2简单,低成本;成像保真度有限细胞、切片观测
    倾斜成像[55]0.69 μm5.6×10−2120 mm2
    编码叠层成像[26]0.3 μm6.7×10−2240 mm2
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  • 收稿日期:  2022-05-13
  • 录用日期:  2022-07-07
  • 网络出版日期:  2022-08-03

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