Open AccessImprovement in focusing accuracy of DNA sequencing microscope with multi-position laser differential confocal autofocus method
Author(s) -
Zhang Xin,
Fei Zeng,
Yang Li,
Yanfeng Qiao
Publication year - 2018
Publication title -
optics express
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.394
H-Index - 271
ISSN - 1094-4087
DOI - 10.1364/oe.26.000887
Subject(s) - autofocus , optics , microscope , focus (optics) , computer science , confocal , laser , position (finance) , computer vision , artificial intelligence , physics , finance , economics
High focusing accuracy in microscopes could improve the imaging quality to reduce the error rate in DNA sequencing. We propose a new feedback method to improve the focusing condition to a very high accuracy. A reference laser reflected by the sample is detected by two or more sensors around the confocal point. After acquiring the signals from the out-of-focus positions, online data processing is implemented to provide feedbacks for real-time focus-plane locking on the sample surface. This method provides an accuracy better than 1/10 of the objective depth-of-focus. To balance optical aberrations, a specific optical feedback system should be designed, with athermal design considerations to adapt DNA sequencing work to temperature fluctuations.