Document Detail


Microtable arrays for culture and isolation of cell colonies.
MedLine Citation:
PMID:  20644916     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Cell microarrays with culture sites composed of individually removable microstructures or micropallets have proven benefits for isolation of cells from a mixed population. The laser energy required to selectively remove these micropallets with attached cells from the array depends on the microstructure surface area in contact with the substrate. Laser energies sufficient to release micropallets greater than 100 μm resulted in loss of cell viability. A new three-dimensional culture site similar in appearance to a table was designed and fabricated using a simple process that relied on a differential sensitivity of two photoresists to UV-mediated photopolymerization. With this design, the larger culture area rests on four small supports to minimize the surface area in contact with the substrate. Microtables up to 250 × 250 μm were consistently released with single 10-μJ pulses to each of the four support structures. In contrast, microstructures with a 150 × 150-μm surface area in contact with the substrate could not be reliably released at pulse energies up to 212 μJ. Cassie-Baxter wetting is required to provide a barrier of air to localize and sequester cells to the culture sites. A second asset of the design was an increased retention of this air barrier under conditions of decreased surface tension and after prolonged culture of cells. The improved air retention was due to the hydrophobic cavity created beneath the table and above the substrate which entrapped air when an aqueous solution was added to the array. The microtables proved an efficient method for isolating colonies from the array with 100% of selected colonies competent to expand following release from the array.
Authors:
Jeng-Hao Pai; Wei Xu; Christopher E Sims; Nancy L Allbritton
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2010-07-20
Journal Detail:
Title:  Analytical and bioanalytical chemistry     Volume:  398     ISSN:  1618-2650     ISO Abbreviation:  Anal Bioanal Chem     Publication Date:  2010 Nov 
Date Detail:
Created Date:  2010-10-28     Completed Date:  2011-02-25     Revised Date:  2011-11-01    
Medline Journal Info:
Nlm Unique ID:  101134327     Medline TA:  Anal Bioanal Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  2595-604     Citation Subset:  IM    
Affiliation:
Department of Chemistry, University of North Carolina, Chapel Hill, NC 27599, USA.
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MeSH Terms
Descriptor/Qualifier:
Cell Culture Techniques / instrumentation,  methods
Cell Separation / instrumentation,  methods*
Culture Media
Equipment Design
Hela Cells
Humans
Microarray Analysis / instrumentation,  methods*
Microscopy, Electron, Scanning
Microscopy, Fluorescence
Surface Properties
Grant Support
ID/Acronym/Agency:
EB007612/EB/NIBIB NIH HHS; HG004843/HG/NHGRI NIH HHS; R01 EB007612-04/EB/NIBIB NIH HHS; R01 HG004843-02/HG/NHGRI NIH HHS
Chemical
Reg. No./Substance:
0/Culture Media

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


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