Biomod/2014/fit Results and Discussion.html: Difference between revisions
Team Fukuoka (talk | contribs) No edit summary |
Team Fukuoka (talk | contribs) No edit summary |
||
| Line 175: | Line 175: | ||
Fig. 8 shows the fluorescence emission spectra measured by microspectroscopy. We used microspectroscopy because the quantity of the sample is very small. The spectrum of FITC-DNA and TAMRA-DNA showed the peaks at 520 nm and 580 nm, respectively, which is the same result as the ones measured on fluorospectrometor (Fig. 8). When the FITC-DNA and TAMRA-DNA are mixed and hybridized, the fluorescence of the FITC is almost quenched while the fluorescence of TAMRA is enhanced, indicating the FRET induced by hybridization. Thus we successfully detect the hybridization by FRET with microspectroscopy. | Fig. 8 shows the fluorescence emission spectra measured by microspectroscopy. We used microspectroscopy because the quantity of the sample is very small. The spectrum of FITC-DNA and TAMRA-DNA showed the peaks at 520 nm and 580 nm, respectively, which is the same result as the ones measured on fluorospectrometor (Fig. 8). When the FITC-DNA and TAMRA-DNA are mixed and hybridized, the fluorescence of the FITC is almost quenched while the fluorescence of TAMRA is enhanced, indicating the FRET induced by hybridization. Thus we successfully detect the hybridization by FRET with microspectroscopy. | ||
<br> | <br> | ||
</p> | </p> | ||
| Line 194: | Line 192: | ||
Fig. 9a and 9b shows the photograph and the SEM image of the amino-functionalized mesoporous silica (MPS-NH2) particle. Spherical particles with the diameter of about 300 nm to 1000 nm are observed. However, because the size of the mesopores is too small, assumed as 2 nm, we could not observed the pores. After the modification with -COOH group (Fig. 9c) and with DNA (Fig. 9d), there were no significant changes in SEM. Therefore, it is understood that the shape of former particle is maintained. | Fig. 9a and 9b shows the photograph and the SEM image of the amino-functionalized mesoporous silica (MPS-NH2) particle. Spherical particles with the diameter of about 300 nm to 1000 nm are observed. However, because the size of the mesopores is too small, assumed as 2 nm, we could not observed the pores. After the modification with -COOH group (Fig. 9c) and with DNA (Fig. 9d), there were no significant changes in SEM. Therefore, it is understood that the shape of former particle is maintained. | ||
<br> | <br> | ||
. | |||
We then observed the samples with CLSM. Fig. 10 shows the CLSM images of the barrel particles, that is the MPS attached with FITC-terminated ssDNA. In Fig. 10, Spherical particles of almost the same size as those observed with SEM is observed. In the fluorescence image at 525±50 nm (Fig. 10b), some green rings are observed, while only very weakly fluorescent image is observed at 595±50 nm (Fig. 10c). Since FITC has the maximum fluorescence wavelength of 525 nm, this result indicate that FITC-DNA was successfully attached to the surface of the mesoporous silica particles. In the combined image (Fig. 10d), the green circles of fluorescence appear to surround the particles. It is thought that, because the ring-like fluorescence is observed, fluorescent molecules with DNA is modified only on the surface of the silica particles. If the fluorescent molecule were fixed also in the pore of inside the particle, and the entire particle should have the fluorescence. | We then observed the samples with CLSM. Fig. 10 shows the CLSM images of the barrel particles, that is the MPS attached with FITC-terminated ssDNA. In Fig. 10, Spherical particles of almost the same size as those observed with SEM is observed. In the fluorescence image at 525±50 nm (Fig. 10b), some green rings are observed, while only very weakly fluorescent image is observed at 595±50 nm (Fig. 10c). Since FITC has the maximum fluorescence wavelength of 525 nm, this result indicate that FITC-DNA was successfully attached to the surface of the mesoporous silica particles. In the combined image (Fig. 10d), the green circles of fluorescence appear to surround the particles. It is thought that, because the ring-like fluorescence is observed, fluorescent molecules with DNA is modified only on the surface of the silica particles. If the fluorescent molecule were fixed also in the pore of inside the particle, and the entire particle should have the fluorescence. | ||
Revision as of 11:55, 25 October 2014
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="ja" lang="ja"> <head> <style type="text/css" >
- dropmenu{
list-style: none; width:900px; height: 55px; margin: 85px auto 100px; padding: 0; background: #8a9b0f; border-bottom: 5px solid #535d09; border-radius: 3px 3px 0 0;
}
- dropmenu li{
position: relative; width: 16.666666666666%; float: left; margin: 0; padding: 0; text-align: center;
}
- dropmenu li a{
display: block; margin: 0; padding: 15px 0 11px; color: #fff; font-size: 16px; font-weight: bold; line-height: 1; text-decoration: none;
}
- dropmenu li:hover > a{
background: #6e7c0c; color: #eff7b1;
}
- dropmenu > li:hover > a{
border-radius: 3px 3px 0 0;
}
- dropmenu li ul{
list-style: none; position: absolute; top: 100%; left: 0; margin: 0; padding:0; border-radius: 0 0 3px 3px;
}
- dropmenu li ul li{
overflow: hidden; width: 100%; height:0; color: #fff; -moz-transition: .2s; -webkit-transition: .2s; -o-transition: .2s; -ms-transition: .2s; transition: .2s;
}
- dropmenu li ul li a{
padding: 13px 15px; background: #6e7c0c; text-align: left; font-size: 16px; font-weight: normal;
}
- dropmenu li:hover ul li{
overflow: visible; height: 43px; border-top: 1px solid #7c8c0e; border-bottom: 1px solid #616d0b;
}
- dropmenu li:hover ul li:first-child{
border-top: 0;
}
- dropmenu li:hover ul li:last-child{
border-bottom: 0;
}
- dropmenu li:hover ul li:last-child a{
border-radius: 0 0 3px 3px;
}
hr {
border-width: 0px 0px 0px 0px; border-style: solid; height: 0px; }
- back {
position: fixed; bottom: 15px; right: 20px;
}
</style>
</head>
<body>
<a name="header"></a> <img src="/images/4/46/Fitaologo.PNG" alt" alt="" width="422" height="98" hspace="0" align="left">
<a name="a"></a>
Preliminary Experiment
verification of FRET with the ssDNAs not modified to the silica
Fig 7 shows the fluorescence spectra of FITC (50 pM), TAMRA-DNA(50 pM), and their mixture. FITC and Tamra-DNA shows fluorescence maxima at 520 and 580 nm, respectively. The mixture of FITC and TAMRA shows just the sum of the spectra of the two dyes, indicating no FRET occurred.
Fig. 8 shows the fluorescence emission spectra measured by microspectroscopy. We used microspectroscopy because the quantity of the sample is very small. The spectrum of FITC-DNA and TAMRA-DNA showed the peaks at 520 nm and 580 nm, respectively, which is the same result as the ones measured on fluorospectrometor (Fig. 8). When the FITC-DNA and TAMRA-DNA are mixed and hybridized, the fluorescence of the FITC is almost quenched while the fluorescence of TAMRA is enhanced, indicating the FRET induced by hybridization. Thus we successfully detect the hybridization by FRET with microspectroscopy.
.
<a name="b"></a> The synthesis of the Barrel particles and Doll particles
Fig. 9a and 9b shows the photograph and the SEM image of the amino-functionalized mesoporous silica (MPS-NH2) particle. Spherical particles with the diameter of about 300 nm to 1000 nm are observed. However, because the size of the mesopores is too small, assumed as 2 nm, we could not observed the pores. After the modification with -COOH group (Fig. 9c) and with DNA (Fig. 9d), there were no significant changes in SEM. Therefore, it is understood that the shape of former particle is maintained.
.
We then observed the samples with CLSM. Fig. 10 shows the CLSM images of the barrel particles, that is the MPS attached with FITC-terminated ssDNA. In Fig. 10, Spherical particles of almost the same size as those observed with SEM is observed. In the fluorescence image at 525±50 nm (Fig. 10b), some green rings are observed, while only very weakly fluorescent image is observed at 595±50 nm (Fig. 10c). Since FITC has the maximum fluorescence wavelength of 525 nm, this result indicate that FITC-DNA was successfully attached to the surface of the mesoporous silica particles. In the combined image (Fig. 10d), the green circles of fluorescence appear to surround the particles. It is thought that, because the ring-like fluorescence is observed, fluorescent molecules with DNA is modified only on the surface of the silica particles. If the fluorescent molecule were fixed also in the pore of inside the particle, and the entire particle should have the fluorescence.
<a name="c"></a>
<a name="d"></a>
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
Abstract
aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
<a name="e"></a>
Abstract
aaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaaa
bbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbbb
| <img alt="" src="/images/0/0b/Fitfisilica.png" width="350" height="350" border="0" /> figure1 |
<img alt="" src="/images/4/4c/Fitkspec.png" width="350" height="350" border="0" /> figure2 |
<img alt="" src="/images/1/16/FitsilicaT.png" width="250" height="250" border="0" /> figure3-1 |
<img alt="" src="/images/4/48/FitsilicaF.png" width="250" height="250" border="0" /> figure3-2 |
<img alt="" src="/images/0/09/FitsilicaTF.png" width="250" height="250" border="0" /> figure3-3 |
</body>
