利用傅立叶平片显微镜对染色体进行高分辨率无油成像。

IF 2 3区 物理与天体物理 Q3 BIOCHEMICAL RESEARCH METHODS
Changliang Yu, Yuguo Tang
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引用次数: 0

摘要

在传统的染色体显微成像中,使用高数值孔径油浸物镜是必不可少的。然而,使用油浸物镜对自动成像系统提出了重大挑战,因为它增加了样品污染和仪器损坏的风险。油浸物镜的浅景深(DOF)也需要更复杂的机械聚焦。本文介绍了一种基于傅立叶显微成像技术的染色体无油显微成像系统。该系统采用100× 0.8 NA干物镜,在524 nm入射波长下实现194 nm的半间距分辨率。在100× 1.25 NA油浸物镜下重建的染色体图像优于常规成像。我们还利用数字调焦方法将有效DOF扩展到±2.4 μm。本研究初步验证了开发新一代染色体扫描仪而不使用油浸物镜的可能性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

High-Resolution Oil-Free Imaging of Chromosomes Using Fourier Ptychographic Microscopy

High-Resolution Oil-Free Imaging of Chromosomes Using Fourier Ptychographic Microscopy

In conventional microscopic imaging of chromosomes, the use of high numerical aperture (NA) oil-immersion objectives is essential. However, the use of oil-immersion objectives poses a significant challenge to automated imaging systems because it increases the risk of sample contamination and instrumental damage. The shallow depth of field (DOF) of oil-immersion objectives also demands more sophisticated mechanical focusing. Here, we introduce a chromosome oil-free microscopic imaging system based on Fourier Ptychographic Microscopy (FPM) technology. The system employs a 100×, 0.8 NA dry objective to achieve a half-pitch resolution of 194 nm at an incident wavelength of 524 nm. The reconstructed images of chromosomes surpassed the conventional imaging with a 100×, 1.25 NA oil-immersion objective. We also utilized digital refocusing methods to extend the effective DOF to ±2.4 μm. This study preliminarily validates the possibility of developing a new generation of chromosome scanners without using an oil-immersion objective.

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来源期刊
Journal of Biophotonics
Journal of Biophotonics 生物-生化研究方法
CiteScore
5.70
自引率
7.10%
发文量
248
审稿时长
1 months
期刊介绍: The first international journal dedicated to publishing reviews and original articles from this exciting field, the Journal of Biophotonics covers the broad range of research on interactions between light and biological material. The journal offers a platform where the physicist communicates with the biologist and where the clinical practitioner learns about the latest tools for the diagnosis of diseases. As such, the journal is highly interdisciplinary, publishing cutting edge research in the fields of life sciences, medicine, physics, chemistry, and engineering. The coverage extends from fundamental research to specific developments, while also including the latest applications.
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