bioimages
Print ISSN : 0919-2719
Regular Article
Comparison of Two Types of Coated Slide Glass Using a New Autofocusing Method for Microscopic Image of Chromosomes
Reiko Kanda, Akira Furukawa, Nobuo Okabe, lsamu Hayata
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ジャーナル フリー

1995 年 3 巻 2 号 p. 101-105

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We recently developed a new autofocusing method for chromosome aberration scoring which enables very fast scanning. This system utilizes, near infrared reflection from the surface of a glass microscope slide coated with an infrared reflector. In practice, another coating over the reflecting film is needed in order to protect the reflecting film from chemical agents. In the present study, the qualities of chromosome preparations on slides coated with SiO2 or SiO over the reflecting film were compared visually and using the automated system. Chromosome preparations were made from cultured human lymphocytes and stained with Giemsa. Microscopic observation showed that stained cytoplasmic substances were found around chromosomes more frequently on SiO2-coated slides than on SiO-coated slides. Metaphases were found, and their images were digitized at high magnification by an automated image analysis system. Cytogeneticists reviewed the digital images and counted false positives and metaphases unsuitable for scoring. “Misframed” metaphases, some of whose chromosomes were straddling the border of the monitor field due to errors of automatic centering or zooming, were observed more frequently on SiO2-coated slides than on SiO-coated slides. The high rate of “misframed” metaphases was a major disadvantage for cytogenetic analysis by automatic facility of image grabbing. Most “misframed” metaphases were irregularly spread, suggesting the interference of SiO2 coating with the circular (uniform) spreading of metaphases. Preparations on SiO-coated glass were superior in terms of the clarity of chromosome images, circular spreading of metaphases, and suitability for automated image analysis.
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