Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
Biol. Res
; 40(1): 29-40, 2007. ilus, graf
Article
em En
| 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.
Palavras-chave
Texto completo:
1
Índice:
LILACS
Assunto principal:
DNA
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Núcleo Celular
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Citometria de Fluxo
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Corantes Fluorescentes
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Indóis
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
Biol Res
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Biol. Res
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Biol. Res. (Online)
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Biological Research
Assunto da revista:
BIOLOGIA
Ano de publicação:
2007
Tipo de documento:
Article
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Project document