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进入三维超分辨:GB-STED
热度 3 xipeng1 2016-1-18 09:36
进入三维超分辨: GB-STED 席鹏 2016-01-14 前面的博文里面我讲过了 STED 这一诺奖技术。该技术的核心是,用一束光制造一个空心光环 ---- 老外叫做 ” 甜甜圈 donut“ (老外咋那么多吃货呢)。有了这个 donut 光,将它套在原来的高斯 PSF 上,利用受激辐射作为擦除机制,即可实现把荧光的 PSF 缩小,也就提高了分辨率。 问题是,要想实现空心光环,特别是要越小越好的空心光环,绝非易事。目前,通常的做法是用一个 0-2 π 的位相调制器,结合圆偏振光进行。这样的话: (1) 对于光学上来说, 2 π 的位相调制就回到了 0 ,因此是一个螺旋式上升的连续结构; (2) 这一位相调制无论从哪个角度剖开,都是一个 0- π 的台阶。 由于从透镜前端到焦点刚好是一个傅里叶变换: 我们可以看到,当 u=0,v=0 时,中心就是对于所有这些 复振幅的积分或求和。由于 , , 这两者刚好相互抵消,因此 中间的极大值变成了极小值,形成了非常美丽的面包圈结构。 图 1 STED 通常 0-2 π 采用涡旋位相光栅实现 donut 。 这一看似美丽的数学解有一个脆弱的地方:对于生物成像,样品会带来新的位相调制。由于生物细胞中,所有细胞器的折射率分布变化大和散射特性不同,导致它们对每一个角度的位相都会产生影响。这个扰动一旦变大, donut 中心就不再为零, STED 将会擦除 PSF 的中心地带,导致分辨率不但不能有效提升,甚至可能会变差。 一句话, donut 不零, STED 不灵。 这也是为什么大家看到很多漂亮的 STED 的成像结果,但都是二维图像的原因。 而另一方面,人们研究了一些抗干扰的光束,如 Bessel 光, Airy 光等,并将其应用在光片成像、 OCT 等领域。 Bessel 光的核心是,通过一个 axicon 对高斯光束进行调制,可以实现一个针状的光分布,且这一针状光斑经过样品时,不会受到样品折射率变化的干扰。 图 2 Bessel 光束调制原理。中心区域通过干涉增强实现了抗干扰的 Bessel 光。 由于 STED 最怕干扰的是中空 donut ,在此条件下,我们把 axicon 和涡旋位相波片结合起来,实现了一个中空的竹子状光束。这一光束同样具有 self healing 的效果。 图 3 Gaussian-Bessel STED 焦点分布示意图。 接下来的工作就豁然开朗了:我们在琼脂样品上测试分辨率,发现在 155 微米的深度,我们能够达到和表面同样的分辨率。我们尝试了折射率失配的 PDMS 样品,发现能够达到 100 微米深度的超分辨。最后,我们制作了一个类脑白质的仿体,发现能够达到 100 微米的穿透深度,实现超分辨。 在过去,很多超分辨的工作受限于样品散射等因素,被局限在二维世界;能研究的样品只有一些细胞,和非常浅层的脑成像。我们希望通过这一技术,实现深层的组织超分辨成像。 相关成果被 Laser Photonics Reviews 作为 2016 年 1 月的封面文章发表。 北京大学物理学院施可彬课题组的于文韬同学为本文第一作者,施可彬研究员和席鹏研究员为共同通讯作者。该工作得到了 973 国家重点基础研究发展计划和国家自然科学基金委的支持。 Wentao Yu et al, Super-resolution deep imaging with hollow Bessel beamSTED microscopy, Laser Photonics Reviews, 10, 147 – 152, 2016. 相关链接: http://onlinelibrary.wiley.com/doi/10.1002/lpor.201500151/abstract
个人分类: STED|11149 次阅读|3 个评论
[转载]MATLAB中的各类贝塞尔函数
nadia1989 2012-11-8 20:16
第一类贝塞尔函数:Jn(x) besselj(nu,Z) computes the Bessel function of the first kind, , for each element of the array Z. The order nu need not be an integer, but must be real. The argument Z can be complex. The result is real where Z is positive. 第二类贝塞尔函数:Yn(x) bessely(nu,Z) computes Bessel functions of the second kind, , for each element of the array Z. The order nu need not be an integer, but must be real. The argument Z can be complex. The result is real where Z is positive. 第三类贝塞尔函数:Hn(x) Bessel function of the third kind (Hankel function) besselh(nu,K,Z) 球 贝塞尔函数: function F = sphericalbessel?(n,x) ifx==0, if n==0, F = 1; else F = 0; end else F=sqrt(pi/(2*x))*bessel?(n+0.5,x); end 其中bessel?代表第一类贝塞尔函数或第二类贝塞尔函数
个人分类: Matlab|4589 次阅读|0 个评论
[转载]很新颖的科普方式
PengJiDing 2012-3-8 14:14
from http://www.st-andrews.ac.uk/seeinglife/science/research/bessel/bessel.html Bessel Beams This is a conversation between Michael, one of our physicists, and James, a member of the public, about the Bessel beam, describing what it is and why it is important for our research! Michael : Hi! I’m a physicist from the University of St Andrews. At the moment we’re very excited about a new type of laser beam that we’ve been doing some research on. It’s called a Bessel beam. James : I see, so what exactly is a Bessel beam? What is it that makes it different? Michael : Well, when you shine a normal laser beam against a wall it shows up as a small point. When you shine a Bessel beam against a wall you get a central dot surrounded by rings of light. Here’s a picture of a Bessel beam. James : I see. What difference do the rings make then? Michael : Well it’s not just the rings. The main difference between a normal laser and a Bessel is the central core. It doesn’t diffract. James : Hmm, I’m not sure I remember what diffraction is. Michael : Diffraction is when light “flares out” after passing through a narrow slit or hole. The thinner the slit the light travels through, the more it flares out. Diffraction doesn’t just happen to light. Ocean waves diffract when they travel through the opening of a harbour wall. Sound diffracts as well. James : So the central core of a Bessel beam doesn’t flare out then? Is that any different to a normal laser beam? They don’t seem to diffract either. Michael : No they don’t seem to diffract but that’s just because of the scale you’re observing them over. You might be able to spot it if you point a laser across a classroom and then put your hand in front. If you were to point a typical laser pen at the moon, then by the time the beam had reached the surface it would have a larger radius than the width of the moon. James : I see, so are we using Bessel beams to point at the moon? That doesn’t seem that useful. Michael : No sorry for the misunderstanding. That’s not what we’re using them for at all. One use we have for them is using them optical injections. James : Optical injections? There’s another phrase I don’t understand. Michael : Don’t worry; it’s definitely not something you would have covered in school. We often want to put new material into a cell. One of the ways we can do this is by using laser light to punch a hole in the cell wall. By placing the cell in a bath of the material we want to insert, the materials should just diffuse in. James : Won’t punching a hole in its wall destroy the cell? Michael : As long as the hole is small enough compared to the cell then it shouldn’t be a problem. Cell walls are actually pretty good at repairing damage to themselves. James : I think I understand optical injections. You use a laser beam like it was a needle to prick the cell wall. Michael : Exactly. James : Well, how does this relate to Bessel beams? Michael : Glad to see you’re paying attention. Well, look at this picture on the right. It shows a close up of a normal laser beam that has been focussed to a point (on top) and a Bessel beam (on the bottom). You can see the difference. James : Yeah. The normal laser beam spreads out hugely compared to the Bessel. Michael : When we’re trying to punch a hole in a cell it’s really important we know exactly how much energy will be hitting the cell. It’s also important to know the area this energy is spread over. James : I think I can see where this is going. With the Bessel beam you could place the cell anywhere along the beam and it would be getting the same energy over the same area. Michael : Exactly. With the normal laser beam it is necessary for the cell to be placed at a very specific place on the beam for it to get the right amount of energy over the right area. This can be quite tricky. Remember, cells are incredibly small. We can fit hundred in just 1 mm. So that means we need to be that precise with our beam. James : Right, well I think I can see how a Bessel beam would make it easier to perform an optical injection but I still have a few more questions. How do we make a Bessel? What about the rings? Do they really never diffract? Michael : I’ll try and take those questions one at a time. Let’s start with how we make a Bessel beam. There are several different methods. The images you can see below are (from left to right) an axicon lens, an annular ring (on the right) and a spatial light modulator (SLM). They all work by altering a normal laser that is either passing through (axicon and the ring) or reflecting off (the SLM). Michael : The details of each method are different but we can visualise the physics of a Bessel beam by thinking about interference patterns of light. James : Remind me how interference works again? Michael : Well, we can think of light acting like a wave. If two light waves meet at a certain point then the result is the two waves added on top of each other. This means that if we get two identical waves meeting when both waves are at peaks then we get a wave with a peak twice as high as the two individual waves. This corresponds to light that is twice as bright. James : I see. So does that mean that if we get two waves meeting when one is at a peak and the other at trough then they cancel each other out and we get a flat line which corresponds to darkness? Michael : Exactly. One final concept to deal with is the idea of extending the wave into all three dimensions. You can see an example to the left. To visualise how a Bessel beam works let’s imagine two extended waves meeting each other travelling at an angle to each other. To the right you can see a top down view of this. The image blue and red lines represent peaks of waves. It is very important to note that the colour in this image is there to make the diagram clearer. In reality both sets of waves would be of identical colour. The two black arrows show us the direction the waves are moving in. We can see that where the two sets of peaks meet we will get positive interference. If we follow the waves back or forward we will begin to build up a line of positive interference. This is represented by the green line and is the central core of the beam. James : So this is how a Bessel beam is made? Michael : Well, first you would have to rotate this image 180˚ out of the page to make a cone of waves but yes, this is one way to think about the physics of a Bessel beam, as a core formed by positive interference. The rings come from further interference. James : About the rings? Do they diffract? What are they useful for? Michael : We don’t use the rings for optical injections but they definitely have other uses. These are in many different fields. The rings do spread out as the beam travels forward but this does not affect the central core. James : This is all beginning to seem a bit too perfect. What about diffraction. Do Bessel beams really never diffract? Michael : A perfect Bessel beam would never diffract however for a perfect Bessel you need infinite energy. This is clearly impossible. So with finite energy we can make approximations of Bessel beams. These approximations to perfect Bessel beams work very well over distances of several cm, which is more than long enough for optical injections. James : Anything else that a Bessel can do that I should be aware of? Michael : Well, there is one more thing. The central core of the Bessel beam can reconstruct itself after being blocked. James : What do you mean by that? Michael : Well, if you were to place an object in the way of a normal laser beam then beyond that object all you would get is shadow. However because a Bessel beam is formed by interference patterns from light travelling at an angle then after a certain distance we get the Bessel central core back again. The gray zone represents the shadow region where we lose the central core. James : This is all a lot to think about! Michael : I agree. There are so many opportunities for Bessel beams beyond the ones you’ve seen today. It can be quite a lot to absorb in one go but if there’s anything you’ve not understood then feel free to get in contact , or better yet come ask us in person at one of our events . This was a contribution from Michael Finlay, one of our undergraduate project students, 2009.
个人分类: 科普|1783 次阅读|0 个评论
贝塞尔函数及其编程和书籍
热度 3 zuozw 2010-5-1 22:49
贝塞尔 函数 ( Bessel function, 又称柱函数 ) 是一类在各工程领域中有着广泛应用的特殊函数。贝塞尔函数是贝塞尔方程 ( 一个二阶常微分方程 ) 的解。 一般有三类贝塞尔函数 1 、第一类贝塞尔函数 (Bessel function of the first kind) J v (x) 2 、第二类贝塞尔函数 (Bessel function of the second kind), 又称诺依曼函数( Neumann function ) :Y v (x)( 或 N v (x)) 3 、第三类贝塞尔函数( Bessel function of the third kind ),又称汉克尔函数( Hankel function ) : H 1 v (x) 和 H 2 v (x) 。 当贝塞尔方程为变形贝塞尔方程 ( 虚宗量贝塞尔方程 ) 时,方程的解一般有两类 1、 变形第一类贝塞尔函数( modified Bessel function of the first kind ) I v (x) 2、 变形第二类贝塞尔函数( modified Bessel function of the second kind ) K v (x) 另外应用比较多的函数还有球贝塞尔函数 (Spherical Bessel functions) 、艾里函数 (Airy functions )等等。由于贝塞尔函数的广泛应用,很多软件都提供了现成的函数供用户使用。 Mathematics :提供了比较多的 Bessel 函数。如: Bessel 函数 BesselJ BesselY BesselI BesselK 球体 Bessel 函数 SphericalBesselJ SphericalBesselY Hankel 函数 HankelH1 HankelH2 SphericalHankelH1 SphericalHankelH2 Airy 函数 AiryAi AiryAiPrime AiryBi AiryBiPrime Kelvin 函数 KelvinBer KelvinBei KelvinKer KelvinKei Struve 函数 StruveH StruveL AngerJ WeberE 零函数 BesselJZero BesselYZero AiryAiZero AiryBiZero Matlab :提供了五个函数 besselj 、 bessely 、 besselh 、 besseli 和 besselk 分别对应第一类 Bessel 函数、第二类 Bessel 函数、第三类 Bessel 函数、第一类变形 Bessel 函数和第二类变形 Bessel 函数。语法 besselj(nu,z) : nu 为函数的阶次,可以不为整数但必须为实数。 z 为函数的变量,可以为复数。还有 besselj(nu,z,1) 语法,可查询帮助。其他函数除了 besselh 有些不同外,基本类似。至于球函数、 Airy 函数等可通过这个函数来构造实现。 Excel :提供了四个函数,即: BESSELJ 、 BESSELY 、 BESSELI 、 BESSELK 。 语法: BESSELJ (x,n) ;其中, x 为参数值, n 为函数的阶数。如果 n 非整数,则截尾取整。即 Excel 中只能计算整数阶次的 Bessel 函数。其他函数类似。 关于贝塞尔函数的专业书籍有: 中文 刘颖,《圆柱函数》国防工业出版社, 1983 。 奚定平,《贝塞尔函数》高等教育出版社和施普林格出版社, 1998. 英文 George Neville Watson 《 A Treatise on the Theory of Bessel Functions 》 Cambridge University Press , 1995(1944 , 1922) 最经典的一本书。 BG Korenev 《 Bessel Functions and Their Applications 》 CRC 2002. Milton Abramowitz, and Irene A. Stegun 《 Handbook of Mathematical Functions with Formulas, Graphs, and Mathematical Tables 》 Dover Publications 1964. I.S. Gradshteyn, I.M. Ryzhik, A Jeffrey, D Zwillinger 《 Table of integrals, series and products 》 (Seventh Edition) Academic Press 2007.
个人分类: 科研心得|12512 次阅读|7 个评论

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