The design of a microscopic system for typical fluorescent in-situ hybridization applications

Dingrong Yi; Shaochuan Xie

doi:10.1117/12.2041743

19 December 2013 The design of a microscopic system for typical fluorescent in-situ hybridization applications

Dingrong Yi, Shaochuan Xie

Proceedings Volume 9045, 2013 International Conference on Optical Instruments and Technology: Optoelectronic Imaging and Processing Technology; 90450O (2013) https://doi.org/10.1117/12.2041743
Event: International Conference on Optical Instruments and Technology (OIT2013), 2013, Beijing, China

Abstract

Fluorescence in situ hybridization (FISH) is a modern molecular biology technique used for the detection of genetic abnormalities in terms of the number and structure of chromosomes and genes. The FISH technique is typically employed for prenatal diagnosis of congenital dementia in the Obstetrics and Genecology department. It is also routinely used to pick up qualifying breast cancer patients that are known to be highly curable by the prescription of Her2 targeted therapy. During the microscopic observation phase, the technician needs to count typically green probe dots and red probe dots contained in a single nucleus and calculate their ratio. This procedure need to be done to over hundreds of nuclei. Successful implementation of FISH tests critically depends on a suitable fluorescent microscope which is primarily imported from overseas due to the complexity of such a system beyond the maturity of the domestic optoelectrical industry. In this paper, the typical requirements of a fluorescent microscope that is suitable for FISH applications are first reviewed. The focus of this paper is on the system design and computational methods of an automatic florescent microscopy with high magnification APO objectives, a fast spinning automatic filter wheel, an automatic shutter, a cooled CCD camera used as a photo-detector, and a software platform for image acquisition, registration, pseudo-color generation, multi-channel fusing and multi-focus fusion. Preliminary results from FISH experiments indicate that this system satisfies routine FISH microscopic observation tasks.

Citation Download Citation

Dingrong Yi and Shaochuan Xie "The design of a microscopic system for typical fluorescent in-situ hybridization applications", Proc. SPIE 9045, 2013 International Conference on Optical Instruments and Technology: Optoelectronic Imaging and Processing Technology, 90450O (19 December 2013); https://doi.org/10.1117/12.2041743

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KEYWORDS

Luminescence

Microscopy

Optical filters

Microscopes

Image filtering

Image registration

Visualization

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