A method for simultaneously delineating multiple t

Authors: F Y Zhao X Yang D Y Chen W Y Ma J G Zheng X M Zhang

Publish Date: 2013/12/04

Volume: 43, Issue: 1, Pages: 53-58

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Abstract

Many studies have suggested a link between the spatial organization of genomes and fundamental biological processes such as genome reprogramming gene expression and differentiation Multicolor fluorescence in situ hybridization on threedimensionally preserved nuclei 3DFISH in combination with confocal microscopy has become an effective technique for analyzing 3D genome structure and spatial patterns of defined nucleus targets including entire chromosome territories and single gene loci This technique usually requires the simultaneous visualization of numerous targets labeled with different colored fluorochromes Thus the number of channels and lasers must be sufficient for the commonly used labeling scheme of 3DFISH “one probeone target” However these channels and lasers are usually restricted by a given microscope system This paper presents a method for simultaneously delineating multiple targets in 3DFISH using limited channels lasers and fluorochromes In contrast to other labeling schemes this method is convenient and simple for multicolor 3DFISH studies which may result in widespread adoption of the technique Lastly as an application of the method the nucleus locations of chromosome territory 18/21 and centromere 18/21/13 in normal human lymphocytes were analyzed which might present evidence of a radial higher order chromatin arrangementThis work was supported by grants from the National Nature Science Foundation of China No 11374180 Chinese National Basic Research Program No 2007CB5119004 Doctoral Program Research Fund of the Chinese Ministry of Education No 20090002110065 and Tsinghua University Initiative Scientific Research Program 2010THZ01

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