Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?

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 a...

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Autores principales:	GALLARDO-ESCÁRATE,CRISTIAN, ÁLVAREZ-BORREGO,JOSUÉ, VON BRAND,ELISABETH, DUPRÉ,ENRIQUE, DEL RÍO-PORTILLA,MIGUEL ÁNGEL
Lenguaje:	English
Publicado:	Sociedad de Biología de Chile 2007
Materias:	Nuclear DNA content fluorescence fading image analysis DAPI
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000100004
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spelling	oai:scielo:S0716-976020070001000042007-11-15Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?GALLARDO-ESCÁRATE,CRISTIANÁLVAREZ-BORREGO,JOSUÉVON BRAND,ELISABETHDUPRÉ,ENRIQUEDEL RÍO-PORTILLA,MIGUEL ÁNGEL Nuclear DNA content fluorescence fading image analysis DAPI 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%. This study demonstrates the feasibility to measure nuclear DNA content by fluorescence fading quantification, as an alternative method concurrently with image analysis proceduresinfo:eu-repo/semantics/openAccessSociedad de Biología de ChileBiological Research v.40 n.1 20072007-01-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000100004en10.4067/S0716-97602007000100004
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	Nuclear DNA content fluorescence fading image analysis DAPI
spellingShingle	Nuclear DNA content fluorescence fading image analysis DAPI GALLARDO-ESCÁRATE,CRISTIAN ÁLVAREZ-BORREGO,JOSUÉ VON BRAND,ELISABETH DUPRÉ,ENRIQUE DEL RÍO-PORTILLA,MIGUEL ÁNGEL Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
description	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%. This study demonstrates the feasibility to measure nuclear DNA content by fluorescence fading quantification, as an alternative method concurrently with image analysis procedures
author	GALLARDO-ESCÁRATE,CRISTIAN ÁLVAREZ-BORREGO,JOSUÉ VON BRAND,ELISABETH DUPRÉ,ENRIQUE DEL RÍO-PORTILLA,MIGUEL ÁNGEL
author_facet	GALLARDO-ESCÁRATE,CRISTIAN ÁLVAREZ-BORREGO,JOSUÉ VON BRAND,ELISABETH DUPRÉ,ENRIQUE DEL RÍO-PORTILLA,MIGUEL ÁNGEL
author_sort	GALLARDO-ESCÁRATE,CRISTIAN
title	Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
title_short	Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
title_full	Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
title_fullStr	Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
title_full_unstemmed	Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?
title_sort	relationship between dapi-fluorescence fading and nuclear dna content: an alternative method to dna quantification?
publisher	Sociedad de Biología de Chile
publishDate	2007
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0716-97602007000100004
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Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?

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