Structural analysis and classification of human metaphase chromosomes by atomic force microscopy

Naoaki Okamoto; Takao Okada

doi:10.1117/12.350628

17 June 1999 Structural analysis and classification of human metaphase chromosomes by atomic force microscopy

Naoaki Okamoto, Takao Okada

Proceedings Volume 3607, Scanning and Force Microscopies for Biomedical Applications; (1999) https://doi.org/10.1117/12.350628
Event: BiOS '99 International Biomedical Optics Symposium, 1999, San Jose, CA, United States

Abstract

We applied atomic force microscopy (AFM) to the analysis and classification of metaphase chromosomes. Human chromosomes were isolated from blood and spread over a glass substrate. We found that air-dried and Giemsa stained chromosomes had a granular surface and the height of approximately 250 nm; however unstained chromosomes had a smooth surface and the height was approximately 100 nm. Giemsa staining caused swelling of the chromosome structure. For the structural analysis, chromosomes were treated with hyaluronidase or a citric acid buffer. The effects of the treatments on chromosomal components, spiral structure and 30-nm solenoid fiber were observed. Each step of G-banding treatments of chromosomes was also visualized by AFM. The trypsin treatment collapsed the chromosomes and subsequent Giemsa staining caused dramatically reswelling of the chromosomes. The height of the G-positive region was approximately 200 nm but the unstained region was approximately 50 nm. The difference in thickness observed was produced by binding of the dye. The AFM image of the banding patterns of treated chromosomes was clearer than the image obtained with an optical microscope. These images made it possible to visualize the karyotyping of chromosomes using AFM. Detection of in situ hybridization using AFM and microdissection of chromosomes using AFM were also investigated.

Citation Download Citation

Naoaki Okamoto and Takao Okada "Structural analysis and classification of human metaphase chromosomes by atomic force microscopy", Proc. SPIE 3607, Scanning and Force Microscopies for Biomedical Applications, (17 June 1999); https://doi.org/10.1117/12.350628

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