DIFFERENTIAL POLARIZATION MICROSCOPY OF CHANGES IN STRUCTURE IN SPERMATOCYTE NUCLEI
DIFFERENTIAL POLARIZATION MICROSCOPY OF CHANGES IN STRUCTURE IN SPERMATOCYTE NUCLEI
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DOI：
10.1038/328452a0
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发表时间：
1987-07-30
期刊：
影响因子：
64.8
通讯作者：
TINOCO, I
中科院分区：
文献类型：
--
作者：
MICKOLS, W;MAESTRE, MF;TINOCO, I
位相相关的显微镜形式(如位相对比显微镜、干涉显微镜或偏振显微镜)已被使用多年；位相延迟的大小主要基于折射率来产生图像。我们已经开发了一种显微镜，它可以根据不同形式的入射偏振光的消光程度的微小差异来形成图像。通过调制入射到样品上的光的偏振，并通过数字记录透射光强度的差异，我们获得了特定地揭示有序线性结构或手性(右手或左手)结构的图像。线偏振光以两个垂直的偏振方向交替入射，形成具有线性顺序或线性取向的结构的图像。右旋和左旋的圆偏振光交替入射到样品上，形成手性结构的图像。既没有线性顺序也没有手性的结构本质上是看不见的。因此，基于线性二向色性、圆二向色性以及线性和圆形差分散射的图像可以用来检测用常规方法可能难以观察到的特定类型的结构。我们用这种“线性和圆形差分成像”来研究果蝇活的初级精母细胞转录活跃或不活跃时核仁(RNA合成的部位)的结构。一些不活跃的核仁是两部分的，通过线偏振光和圆偏振光的差示散射可以看到两种不同的结构。活性核仁是一个单一的结构域，它与Y染色体的一部分有明显的区别，在线偏振光和圆偏振光的作用下，它显示出不同的内部结构。因此，偏振相关的图像揭示了与细胞转录活动相关的结构。
Phase-dependent forms of microscopy (such as phase contrast, interference, or polarization microscopy) have been used for many years; the amount of phase retardation produces images based mainly on index of refraction. We have developed a microscope that forms images that depend on small differences in extinction for different forms of incident polarized light. By modulating the polarization of light incident on the sample and digitally recording the difference in intensities of transmitted light, we obtain images which specifically reveal either ordered linear structures or chiral (right- or left-handed) structures. Linearly polarized light, incident alternately with two perpendicular directions of polarization, forms images of structures which have linear order or linear orientation. Right-handed and left-handed circularly polarized light incident alternately on a sample forms images of chiral structures. Structures with neither linear order nor chirality are essentially invisible. Thus, images based on linear dichroism, circular dichroism, and linear and circular differential scattering can be used to detect specific types of structures which may be difficult to observe by conventional methods. We have used such 'linear and circular differential imaging' to study the structure of the nucleolus (the site of RNA synthesis) in live primary spermatocytes of Drosophila when they are transcriptionally active or inactive. Some inactive nucleoli are bipartite, with two distinct structures visible by differential scattering of both linearly and circularly polarized light. The active nucleolus is a single domain; it is clearly distinguished from part of the Y chromosome, and it shows different internal structure with linearly and circularly polarized light. Thus, polarization-dependent images reveal structures which can be associated with the transcriptional activity of cells.