Rapid Autofocus Method Based on LED Oblique Illumination for Metaphase Chromosome Microscopy Imaging System
Year: 2024
Type: article
Abstract: In clinical practice, microscopes are commonly used for imaging chromosomes to diagnose genetic diseases. Achieving precise and rapid autofocusing is a significant challenge in the advancement of high-throughput chromosome imaging systems. Here, we introduce a rapid autofocus method based on LED oblique illumination for dual-objective configuration in metaphase chromosome imaging system. Our method utilizes a programmable LED array for sample illumination, employing a sequential activation of two LEDs from opposing angles to create oblique illumination. The defocus distance is ascertained through image cross-correlation calculation. Illumination with multiple LEDs in the array is used to acquire bright-field images after completing the focusing. Our method can perform continuous autofocus under a 10× objective and a 100× oil immersion objective, with average focusing errors of 1.29 μm and 0.12 μm, respectively. The total imaging time for a single sample has been significantly reduced from approximately 10 min with conventional methods to just 2 min. This study provides preliminary evidence supporting the viability of developing a next-generation high-throughput chromosome scanner employing a LED array.
Source: Photonics
Authors: Changliang Yu, Fangqiu Ding, Zhenyu Ma, Yuguo Tang
Language: English
Subfield: Media Technology
Field: Engineering
Domain: Physical Sciences
Open Access status: gold
Estimated APC paid: $2,728
OpenAlex ID: W4404536880