Abstract
Background: Optimizing operational parameters of the digital microscope system is an important technique to acquire high quality cytogenetic images and facilitate the process of karyotyping so that the efficiency and accuracy of diagnosis can be improved.OBJECTIVE: This study investigated the impact of the condenser on cytogenetic image quality and system working performance using a prototype digital microscope image scanning system.Methods: Both theoretical analysis and experimental validations through objectively evaluating a resolution test chart and subjectively observing large numbers of specimen were conducted.Results: The results show that the optimal image quality and large depth of field (DOF) are simultaneously obtained when the numerical aperture of condenser is set as 60%–70% of the corresponding objective. Under this condition, more analyzable chromosomes and diagnostic information are obtained. As a result, the system shows higher working stability and less restriction for the implementation of algorithms such as autofocusing especially when the system is designed to achieve high throughput continuous image scanning.Conclusions: Although the above quantitative results were obtained using a specific prototype system under the experimental conditions reported in this paper, the presented evaluation methodologies can provide valuable guidelines for optimizing operational parameters in cytogenetic imaging using the high throughput continuous scanning microscopes in clinical practice.
Highlights
Karyotyping of human chromosomes based on microscopic cytogenetic imaging has been routinely performed in cytogenetic laboratories as a standard procedure in diagnosis of cancers and genetic disorders through the analysis of chromosome pattern changes and/or aberration [1,2,3]
As one of the most significant components in the optical microscopes, the condenser controls the illumination intensity and the angle of light entering the front lens of the objective, and impacts the image sharpness, resolution, and depth of field (DOF) and the imaging quality of the whole system
Understanding the condenser impacts and finding out the optimal relationship between the condenser and the objective based on the theoretical analysis and the comprehensively experimental validations is a critical issue for optimizing operational parameters of digital optical microscopes
Summary
Karyotyping of human chromosomes based on microscopic cytogenetic imaging has been routinely performed in cytogenetic laboratories as a standard procedure in diagnosis of cancers and genetic disorders through the analysis of chromosome pattern changes and/or aberration [1,2,3]. Acquiring cytogenetic images with sufficient band pattern sharpness and high resolution is important and helpful to improve disease diagnosis accuracy and efficiency in analyzing automatically scanned and acquired digital microscopic images [4]. Ren et al / The impact of the condenser on cytogenetic image quality systems and the computer aided detection (CAD) schemes to assist selection of analyzable metaphase chromosomes and conduct related image process tasks in recent years [5,6,7] Among these efforts, optimizing operational parameters becomes a critical issue to achieve the accurate and reliable process of high throughput and continuous image scanning, which will help improve the efficiency of chromosome identification and classification. CONCLUSIONS: the above quantitative results were obtained using a specific prototype system under the experimental conditions reported in this paper, the presented evaluation methodologies can provide valuable guidelines for optimizing operational parameters in cytogenetic imaging using the high throughput continuous scanning microscopes in clinical practice
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