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Micromachines 2014, 5(2), 158-170; doi:10.3390/mi5020158

A Semi-Closed Device for Chromosome Spreading for Cytogenetic Analysis

1
Department of Micro- and Nanotechnology, Technical University of Denmark, Orsteds Plads, 2800 Kgs. Lyngby 2800, Denmark
2
Center for Molecular Biology 'Severo Ochoa', UAM campus Cantoblano, Madrid 28049, Spain
3
Faculty of Health Sciences, Department of Cellular and Molecular Medicine, University of Copenhagen, Copenhagen 2200, Denmark
4
Applied Human Molecular Genetics, Kennedy Center, Copenhagen University Hospital, Rigshospitalet, Glostrup 2600, Denmark
*
Author to whom correspondence should be addressed.
Received: 3 February 2014 / Revised: 27 March 2014 / Accepted: 28 March 2014 / Published: 3 April 2014
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Abstract

Metaphase chromosome spreading is the most crucial step required for successful karyotyping and FISH analysis. These two techniques are routinely used in cytogenetics to assess the chromosome abnormalities. The spreading process has been studied for years but it is still considered an art more than a science. The chromosome spreading greatly depends on the environmental conditions such as humidity and temperature, which govern the evaporation of fixative, in which the cells are suspended. The spreading is normally performed manually in ambient conditions on glass slides, which are hydrophilic, and thus allow for better quality spreads. Further cytogenetic analysis depends on the quality of the spreads, which is dependent on the skills of the personnel and is thus limited to laboratory settings. Here, we present a semi-closed microfluidic chip for preparation of the metaphase spreads on a glass and a Topasr substrate rendered more hydrophilic by oxygen plasma treatment coupled with photografting. The device consists of a microfluidic chamber with perfusion holes that facilitate the evaporation of fixative and reliable formation of the spreads. The usability of the chromosome spreads formed on the glass and the Topasr slide is tested by performing FISH analysis.
Keywords: cytogenetic analysis; evaporation; chromosome spreading; microfluidic chip cytogenetic analysis; evaporation; chromosome spreading; microfluidic chip
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

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MDPI and ACS Style

Kwasny, D.; Mednova, O.; Vedarethinam, I.; Dimaki, M.; Silahtaroglu, A.; Tümer, Z.; Almdal, K.; Svendsen, W.E. A Semi-Closed Device for Chromosome Spreading for Cytogenetic Analysis. Micromachines 2014, 5, 158-170.

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