Document Detail


Photoporation and cell transfection using a violet diode laser.
MedLine Citation:
PMID:  19488389     Owner:  NLM     Status:  In-Data-Review    
Abstract/OtherAbstract:
The introduction and subsequent expression of foreign DNA inside living mammalian cells (transfection) is achieved by photoporation with a violet diode laser. We direct a compact 405 nm laser diode source into an inverted optical microscope configuration and expose cells to 0.3 mW for 40 ms. The localized optical power density of ~1200 MW/m2 is six orders of magnitude lower than that used in femtosecond photoporation (~104 TW/m2). The beam perforates the cell plasma membrane to allow uptake of plasmid DNA containing an antibiotic resistant gene as well as the green fluorescent protein (GFP) gene. Successfully transfected cells then expand into clonal groups which are used to create stable cell lines. The use of the violet diode laser offers a new and simple poration technique compatible with standard microscopes and is the simplest method of laser-assisted cell poration reported to date.
Authors:
L Paterson; B Agate; M Comrie; R Ferguson; T Lake; J Morris; A Carruthers; C T Brown; W Sibbett; P Bryant; F Gunn-Moore; A Riches; Kishan Dholakia
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Optics express     Volume:  13     ISSN:  1094-4087     ISO Abbreviation:  Opt Express     Publication Date:  2005 Jan 
Date Detail:
Created Date:  2009-06-02     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101137103     Medline TA:  Opt Express     Country:  United States    
Other Details:
Languages:  eng     Pagination:  595-600     Citation Subset:  -    
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