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Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
Gallardo-Escárate, Cristian; Álvarez-Borrego, Josué; Von Brand, Elisabeth; Dupré, Enrique; Del Río-Portilla, Miguel Angel.
Afiliação
  • Gallardo-Escárate, Cristian; Universidad de Concepción. Facultad de Ciencias Naturales y Oceanógraficas. Departamento de Oceanografía. Concepción. CL
  • Álvarez-Borrego, Josué; Centro de Investigación Científica y de Educación Superior de Ensenada. Departamento de Optica. Ensenada. MX
  • Von Brand, Elisabeth; Universidad Católica del Norte. Departamento de Biología Marina. Coquimbo. CL
  • Dupré, Enrique; Universidad Católica del Norte. Departamento de Biología Marina. Coquimbo. CL
  • Del Río-Portilla, Miguel Angel; Centro de Investigación Científica y de Educación Superior de Ensenada. Departamento de Acuicultura. Ensenada. MX
Biol. Res ; 40(1): 29-40, 2007. ilus, graf
Artigo em Inglês | LILACS | ID: lil-456606
Biblioteca responsável: BR1.1
ABSTRACT
In observations by confocal or conventional fluorescence microscopy, important factors should be considered in order to obtain accurate images. One of them, such as the fluorescence bleaching from highest intensity to lowest signal of fluorescence is a common problem with several DNA fluorochromes and especially for DAPI stain. The fluorescence of DAPI fades rapidly when it is exposed to UV light, under optimal conditions of observation. Although the fading process can be retarded using a mounting medium with antifading reagents, the photochemical process underlying the fluorescence decay has not yet been fully explained. In addition, no relationship between fluorescence fading and nuclear DNA content has been tested. In order to test this relationship, we measured by means of image analysis the DAPI-fluorescence intensity in several cellular types (spermatozoa, erythrocytes and haemocytes) during their fluorescence bleaching. An algorithm specifically built in MATLAB software was used for this approach. The correlation coefficient between nuclear DNA content and DAPI-fluorescence fading was found equal to 99 percent. This study demonstrates the feasibility to measure nuclear DNA content by fluorescence fading quantification, as an alternative method concurrently with image analysis procedures.
Assuntos
Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: LILACS Assunto principal: DNA / Núcleo Celular / Citometria de Fluxo / Corantes Fluorescentes / Indóis Limite: Animais Idioma: Inglês Revista: Biol. Res Assunto da revista: Biologia Ano de publicação: 2007 Tipo de documento: Artigo / Documento de projeto País de afiliação: Chile / México Instituição/País de afiliação: Centro de Investigación Científica y de Educación Superior de Ensenada/MX / Universidad Católica del Norte/CL / Universidad de Concepción/CL

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Texto completo: Disponível Coleções: Bases de dados internacionais Base de dados: LILACS Assunto principal: DNA / Núcleo Celular / Citometria de Fluxo / Corantes Fluorescentes / Indóis Limite: Animais Idioma: Inglês Revista: Biol. Res Assunto da revista: Biologia Ano de publicação: 2007 Tipo de documento: Artigo / Documento de projeto País de afiliação: Chile / México Instituição/País de afiliação: Centro de Investigación Científica y de Educación Superior de Ensenada/MX / Universidad Católica del Norte/CL / Universidad de Concepción/CL
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