Document Detail


Towards a cellular multi-parameter analysis platform: fluorescence in situ hybridization (FISH) on microhole-array chips.
MedLine Citation:
PMID:  22256298     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Highly-sensitive analysis systems based on cellular multi-parameter are needed in the diagnostics. Therefore we improved our previously developed chip platform for another additional analysis method, the fluorescence in situ hybridization. Fluorescence in situ hybridization (FISH) is a technique used in the diagnostics to determine the localization and the presence or absence of specific DNA sequence. To improve this labor- and cost-intensive method, we reduced the assay consumption by a factor of 5 compared to the standard protocol. Microhole chips were used for making the cells well addressable. The chips were fabricated by semiconductor technology on the basis of a Silicon wafer with a thin deposited silicon nitride layer (Si(3)N(4)). Human retina pigment epithelia (ARPE-19) cells were arrayed on 5-μm holes of a 35 × 35 microhole-array by a gently negative differential pressure of around 5 mbar. After 3 hours of incubation the cells were attached to the chip and the FISH protocol was applied to the positioned cells. A LabView software was developed to simplify the analysis. The software automatically counts the number of dots (positive labeled chromosome regions) as well as the distance between adjacent dots. Our developed platform reduces the assay consumption and the labor time. Furthermore, during the 3 hours of incubation non-invasive or minimal-invasive methods like Raman- and impedance-spectroscopy can be applied.
Authors:
Christian M Kurz; Stefan V D Moosdijk; Hagen Thielecke; Thomas Velten
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Conference proceedings : ... Annual International Conference of the IEEE Engineering in Medicine and Biology Society. IEEE Engineering in Medicine and Biology Society. Annual Conference     Volume:  2011     ISSN:  1557-170X     ISO Abbreviation:  Conf Proc IEEE Eng Med Biol Soc     Publication Date:  2011  
Date Detail:
Created Date:  2012-01-18     Completed Date:  2012-05-31     Revised Date:  2014-08-21    
Medline Journal Info:
Nlm Unique ID:  101243413     Medline TA:  Conf Proc IEEE Eng Med Biol Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  8408-11     Citation Subset:  IM    
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MeSH Terms
Descriptor/Qualifier:
Cell Line
Fluorescein-5-isothiocyanate / chemistry
Humans
Image Processing, Computer-Assisted
In Situ Hybridization, Fluorescence / methods*
Light
Microfluidic Analytical Techniques / methods*
Microscopy, Fluorescence
Software
Xanthenes / chemistry
Chemical
Reg. No./Substance:
0/Xanthenes; 82354-19-6/Texas red; I223NX31W9/Fluorescein-5-isothiocyanate

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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