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Inorganic phosphate nanorods are a novel fluorescent label in cell biology.

Patra CR, Bhattacharya R, Patra S, Basu S, Mukherjee P, Mukhopadhyay D - J Nanobiotechnology (2006)

Bottom Line: We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology.These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC).The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic Cancer Center, Mayo Clinic, Rochester, Minnesota, USA. patra.chittaranjan@mayo.edu

ABSTRACT
We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology. These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC). The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM). At concentrations up to 50 microg/ml, the use of [3H]-thymidine incorporation assays, apoptosis assays (TUNEL), and trypan blue exclusion illustrated the non-toxic nature of these nanorods, a major advantage over traditional organic dyes.

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Emission spectra of (A) EuPO4·H2O nanorods loaded inside 786-O cells treated at various concentrations (a = 0 μg/ml, b = 50 μg/ml, c = 100 μg/ml), (B) TbPO4·H2O nanorods loaded inside HUVEC cells treated at various concentrations (a = 0 μg/ml, b = 20 μg/ml, c = 50 μg/ml, d = 100 μg/ml).
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Figure 3: Emission spectra of (A) EuPO4·H2O nanorods loaded inside 786-O cells treated at various concentrations (a = 0 μg/ml, b = 50 μg/ml, c = 100 μg/ml), (B) TbPO4·H2O nanorods loaded inside HUVEC cells treated at various concentrations (a = 0 μg/ml, b = 20 μg/ml, c = 50 μg/ml, d = 100 μg/ml).

Mentions: In order to determine if the fluorescence activity of these LnPO4·H2O nanorods remain unchanged inside the cell, 786-O cells and HUVEC are incubated for 24 hours with these nanorods at various concentrations and the emission (fluorescence) spectra were recorded on a Fluorolog-3 Spectrofluorometer after extensive washing with PBS (phosphate buffer saline) and shown in Figure 3A–B. Figure 3A shows the emission spectra of 786-O cells loaded with EuPO4·H2O nanorods at different concentrations: 0 μg/ml (curve-a), 50 μg/ml (curve-b), and 100 μg/ml (curve-c), respectively. Similarly, Figure 3B shows the emission spectra of HUVEC cells loaded with TbPO4·H2O nanorods at different concentrations: 0 μg/ml (curve-a), 20 μg/ml (curve-b), 50 μg/ml (curve-c), and 100 μg/ml (curve-d), respectively. Similar results were obtained when 786-O cells were treated with TbPO4·H2O and HUVEC cells were treated with EuPO4·H2O nanorods (data not shown). It was observed that with increasing concentrations of LnPO4·H2O nanorods (0 to 100 μg/ml), the rate of nanorod accumulation inside the 786-O and HUVEC cells increased as the fluorescence intensity from curve -a to curve -c/d increased (Figure 3A–B). As these nanorods show their distinct fluorescence properties inside the HUVEC and 786-O cells, it indirectly proves that these nanorods are internalized (which is confirmed by TEM, as discussed later).


Inorganic phosphate nanorods are a novel fluorescent label in cell biology.

Patra CR, Bhattacharya R, Patra S, Basu S, Mukherjee P, Mukhopadhyay D - J Nanobiotechnology (2006)

Emission spectra of (A) EuPO4·H2O nanorods loaded inside 786-O cells treated at various concentrations (a = 0 μg/ml, b = 50 μg/ml, c = 100 μg/ml), (B) TbPO4·H2O nanorods loaded inside HUVEC cells treated at various concentrations (a = 0 μg/ml, b = 20 μg/ml, c = 50 μg/ml, d = 100 μg/ml).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC1636656&req=5

Figure 3: Emission spectra of (A) EuPO4·H2O nanorods loaded inside 786-O cells treated at various concentrations (a = 0 μg/ml, b = 50 μg/ml, c = 100 μg/ml), (B) TbPO4·H2O nanorods loaded inside HUVEC cells treated at various concentrations (a = 0 μg/ml, b = 20 μg/ml, c = 50 μg/ml, d = 100 μg/ml).
Mentions: In order to determine if the fluorescence activity of these LnPO4·H2O nanorods remain unchanged inside the cell, 786-O cells and HUVEC are incubated for 24 hours with these nanorods at various concentrations and the emission (fluorescence) spectra were recorded on a Fluorolog-3 Spectrofluorometer after extensive washing with PBS (phosphate buffer saline) and shown in Figure 3A–B. Figure 3A shows the emission spectra of 786-O cells loaded with EuPO4·H2O nanorods at different concentrations: 0 μg/ml (curve-a), 50 μg/ml (curve-b), and 100 μg/ml (curve-c), respectively. Similarly, Figure 3B shows the emission spectra of HUVEC cells loaded with TbPO4·H2O nanorods at different concentrations: 0 μg/ml (curve-a), 20 μg/ml (curve-b), 50 μg/ml (curve-c), and 100 μg/ml (curve-d), respectively. Similar results were obtained when 786-O cells were treated with TbPO4·H2O and HUVEC cells were treated with EuPO4·H2O nanorods (data not shown). It was observed that with increasing concentrations of LnPO4·H2O nanorods (0 to 100 μg/ml), the rate of nanorod accumulation inside the 786-O and HUVEC cells increased as the fluorescence intensity from curve -a to curve -c/d increased (Figure 3A–B). As these nanorods show their distinct fluorescence properties inside the HUVEC and 786-O cells, it indirectly proves that these nanorods are internalized (which is confirmed by TEM, as discussed later).

Bottom Line: We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology.These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC).The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM).

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Mayo Clinic Cancer Center, Mayo Clinic, Rochester, Minnesota, USA. patra.chittaranjan@mayo.edu

ABSTRACT
We report the first use of inorganic fluorescent lanthanide (europium and terbium) ortho phosphate [LnPO4.H2O, Ln = Eu and Tb] nanorods as a novel fluorescent label in cell biology. These nanorods, synthesized by the microwave technique, retain their fluorescent properties after internalization into human umbilical vein endothelial cells (HUVEC), 786-O cells, or renal carcinoma cells (RCC). The cellular internalization of these nanorods and their fluorescence properties were characterized by fluorescence spectroscopy (FS), differential interference contrast (DIC) microscopy, confocal microscopy, and transmission electron microscopy (TEM). At concentrations up to 50 microg/ml, the use of [3H]-thymidine incorporation assays, apoptosis assays (TUNEL), and trypan blue exclusion illustrated the non-toxic nature of these nanorods, a major advantage over traditional organic dyes.

No MeSH data available.


Related in: MedlinePlus