Carboxy-eosin As A Marker For Correlative Light - Electron Microscopic Imaging Of Newly Synthesized In Vivo DNA

Alexandre Loukanova,c*, Saim  Eminb, Surya  P. Singhb, Anatoli  Angelova and Kuniaki Nagayamac

aLaboratory of Eng. Nanobiotechnology, Department of Engineering Geoecology, University of Mining and Geology, Sofia 1700, Bulgaria

bNational Institute for Materials Science, Advanced Photovoltaics Cente, Tsukuba 3050047, Japan

cDivision of Nano-Structure Physiology, Okazaki Institute for Integrative Bioscience, Myodaiji-cho, Okazaki 4448787, Japan

Adv. Mater. Lett., 2010, 1 (2), pp 114-117

DOI: 10.5185/amlett.2010.5127

Publication Date (Web): Apr 08, 2012

E-mail: alex_loukanov@yahoo.com

Abstract


Here we report carboxy-eosin as a fluorescent marker for in vivo visualization of DNA in the correlative (fluorescence and electron) biological microscope. For that purpose a fluorescent thymidine analog 2’,4’,5’,7’-tetrabromofluorescein-aminoallyl-dUTP (eosin-aminoallyl-dUTP) was synthesized and incorporated into the genome of Escherichia coli. The fluorescent dye eosin contains four bromine atoms, which enable electron spectroscopic imaging in the transmission electron microscope. Labeled cells were prepared and directly observed by fluorescence and electron microscope without any preliminary fixation step. We demonstrated that eosin-aminoallyl-dUTP is a suitable marker for imaging of cell’s ultrastructures by correlative biological microscopy.

Keywords

Carboxy-eosin, correlative microscopy, Hilbert differential contrast TEM, ESI

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