Center for Life Nano Science@Sapienza, Istituto Italiano di Tecnologia, Rome, Italy.
Department of Physics, University of Rome "Sapienza", Rome, Italy.
Sci Rep. 2017 Feb 7;7(1):17. doi: 10.1038/s41598-017-00042-w.
An ideal direct imaging system entails a method to illuminate on command a single diffraction-limited region in a generally thick and turbid volume. The best approximation to this is the use of large-aperture lenses that focus light into a spot. This strategy fails for regions that are embedded deep into the sample, where diffraction and scattering prevail. Airy beams and Bessel beams are solutions of the Helmholtz Equation that are both non-diffracting and self-healing, features that make them naturally able to outdo the effects of distance into the volume but intrinsically do not allow resolution along the propagation axis. Here, we demonstrate diffraction-free self-healing three-dimensional monochromatic light spots able to penetrate deep into the volume of a sample, resist against deflection in turbid environments, and offer axial resolution comparable to that of Gaussian beams. The fields, formed from coherent mixtures of Bessel beams, manifest a more than ten-fold increase in their undistorted penetration, even in turbid milk solutions, compared to diffraction-limited beams. In a fluorescence imaging scheme, we find a ten-fold increase in image contrast compared to diffraction-limited illuminations, and a constant axial resolution even after four Rayleigh lengths. Results pave the way to new opportunities in three-dimensional microscopy.
理想的直接成像系统需要有一种方法,能够按需照亮通常较厚且混浊的体积中的单个衍射受限区域。最接近这种方法的是使用大孔径透镜将光聚焦成一个点。对于嵌入到样本深处的区域,这种策略会失败,因为在这些区域中,衍射和散射占主导地位。艾里光束和贝塞尔光束是亥姆霍兹方程的解,它们都具有非衍射和自修复的特性,这些特性使它们能够自然地克服进入体积的距离的影响,但本质上不允许沿着传播轴进行分辨率。在这里,我们展示了无衍射自修复的三维单色光斑,能够穿透样本的深层,在混浊环境中抵抗偏转,并提供与高斯光束相当的轴向分辨率。由贝塞尔光束相干混合物形成的场,即使在混浊的牛奶溶液中,与衍射受限光束相比,其未失真的穿透率也增加了十倍以上。在荧光成像方案中,我们发现与衍射受限照明相比,图像对比度提高了十倍,即使在四个瑞利长度之后,轴向分辨率仍然保持不变。这些结果为三维显微镜开辟了新的机会。