利用傅里叶叠层术显微镜实现高分辨率染色体成像：初步评估。

Using Fourier ptychography microscopy to achieve high-resolution chromosome imaging: an initial evaluation.

机构信息

University of Oklahoma, Stephenson School of Biomedical Engineering, Norman, Oklahoma, United States.

University of Oklahoma, School of Electrical and Computer Engineering, Norman, Oklahoma, United States.

出版信息

J Biomed Opt. 2022 Jan;27(1). doi: 10.1117/1.JBO.27.1.016504.

DOI:10.1117/1.JBO.27.1.016504

PMID:35102727

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC8802907/

Abstract

SIGNIFICANCE

Searching analyzable metaphase chromosomes is a critical step for the diagnosis and treatment of leukemia patients, and the searching efficiency is limited by the difficulty that the conventional microscopic systems have in simultaneously achieving high resolution and a large field of view (FOV). However, this challenge can be addressed by Fourier ptychography microscopy (FPM) technology.

AIM

The purpose of this study is to investigate the feasibility of utilizing FPM to reconstruct high-resolution chromosome images.

APPROACH

An experimental FPM prototype, which was equipped with 4 × / 0.1 NA or 10 × / 0.25 NA objective lenses to achieve a theoretical equivalent NA of 0.48 and 0.63, respectively, was developed. Under these configurations, we first generated the system modulation transfer function (MTF) curves to assess the resolving power. Next, a group of analyzable metaphase chromosomes were imaged by the FPM system, which were acquired from the peripheral blood samples of the leukemia patients. The chromosome feature qualities were evaluated and compared with the results accomplished by the corresponding conventional microscopes.

RESULTS

The MTF curve results indicate that the resolving power of the 4 × / 0.1 NA FPM system is equivalent and comparable to the 20 × / 0.4 NA conventional microscope, whereas the performance of the 10 × / 0.25 NA FPM system is close to the 60 × / 0.95 NA conventional microscope. When imaging the chromosomes, the feature qualities of the 4 × / 0.1 NA FPM system are comparable to the results under the conventional 20 × / 0.4 NA lens, whereas the feature qualities of the 10 × / 0.25 NA FPM system are better than the conventional 60 × / 0.95 NA lens and comparable to the conventional 100 × / 1.25 NA lens.

CONCLUSIONS

This study initially verified that it is feasible to utilize FPM to develop a high-resolution and wide-field chromosome sample scanner.

摘要

意义

寻找可分析的中期染色体是白血病患者诊断和治疗的关键步骤，而传统的显微镜系统在同时实现高分辨率和大视场 (FOV) 方面存在困难，这限制了搜索效率。然而，傅里叶相衬显微镜 (FPM) 技术可以解决这一挑战。

目的

本研究旨在探讨利用 FPM 重建高分辨率染色体图像的可行性。

方法

我们开发了一个实验性的 FPM 原型，该原型配备了 4×/0.1 NA 或 10×/0.25 NA 物镜，理论等效数值孔径分别为 0.48 和 0.63。在这些配置下，我们首先生成系统调制传递函数 (MTF) 曲线来评估分辨率。接下来，我们使用 FPM 系统对来自白血病患者外周血样本的一组可分析中期染色体进行成像。评估染色体特征质量，并将其与相应的传统显微镜的结果进行比较。

结果

MTF 曲线结果表明，4×/0.1 NA FPM 系统的分辨率与 20×/0.4 NA 传统显微镜相当且可相媲美，而 10×/0.25 NA FPM 系统的性能接近 60×/0.95 NA 传统显微镜。在对染色体进行成像时，4×/0.1 NA FPM 系统的特征质量与传统 20×/0.4 NA 镜头下的结果相当，而 10×/0.25 NA FPM 系统的特征质量优于传统 60×/0.95 NA 镜头，与传统 100×/0.125 NA 镜头相当。