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一种基于 DAPI 的改良 C-带技术,可快速实现染色体着丝粒的高摄影对比度。

A DAPI-Based Modified C-banding Technique for a Rapid Achieving High Photographic Contrast of Centromeres on Chromosomes.

机构信息

Nuclear Research Center Negev, 84190, Beer-Sheba, Israel.

Department of Biomedical Engineering, Ben Gurion University, Beer Sheva, Israel.

出版信息

Cell Biochem Biophys. 2022 Jun;80(2):375-384. doi: 10.1007/s12013-022-01065-5. Epub 2022 Feb 8.

Abstract

Many chromosome assays rely on the quantification of chromosome abnormalities in cells, and one important abnormality is the existence of more than one centromere for each chromosome. The quantification of such abnormalities has been studied before. However, this process is labor-intensive and time consuming. Thus, this assay is challenging for ex-laboratory applications, where speed is required. We present a visualization method that uses a cheap stain-DAPI, long (e.g., high-resolution) chromosomes and our modified C-banding method for labeling chromosomes. The labeled chromosomes can then be easily seen with a conventional and readily available fluorescence microscopy system. This method achieves an acceleration of the detection of the presence of constitutive heterochromatin in chromosomal centromeres by more than 10 times, to ~2 h, in Human lymphocyte cells and in cells of the human Jurkat line. This new procedure will ultimately provide an easier and cheaper alternative to FISH/PNA probes, or the classic Giemsa staining method. Simplification and reduction in time of the overall procedure will enable the utilization of centromere-counting assays in laboratory and ex-laboratory applications, including in emergency response.

摘要

许多染色体检测依赖于细胞染色体异常数量的定量分析,其中一个重要的异常是每条染色体存在多个着丝粒。此类异常的定量分析此前已有研究。然而,该过程既耗时又费力。因此,对于需要速度的实验室外应用来说,该检测极具挑战性。我们提出了一种可视化方法,该方法使用廉价的 DAPI 染色剂、长(例如,高分辨率)染色体和我们改进的 C-带染色方法来标记染色体。然后可以使用常规且易于获得的荧光显微镜系统轻松观察标记的染色体。该方法使人类淋巴细胞和人 Jurkat 系细胞中染色体着丝粒的组成型异染色质的存在检测速度提高了 10 倍以上,达到~2 小时。该新程序最终将为 FISH/PNA 探针或经典的吉姆萨染色方法提供更简单、更廉价的替代方法。整个过程的简化和时间的减少将使着丝粒计数检测能够在实验室和实验室外应用中使用,包括在应急响应中使用。

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