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<hr>
<div id="header"><h1><p style="background:#CCFFFF; color:purple;"><font face=cursive size="6"><B>  Team FIT </font> </B></p></a></h1></div>
<br>
         


<table> <tr align="center">
<td cellspacing="10" cellpadding="10" width="1055" width="180" height="60" bgcolor="#BAD3FF"><a href="http://openwetware.org/wiki/Biomod/2014/Fukuoka"><B><font face=cursive color="#003366" size="3">Top</font></B></a></td>
<td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Introduction.html"><B><font face=cursive color="#003366" size="3">Introduction</font></B></a></td>
<td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Approach and Goals.html"><B><font face=cursive  color="#003366" size="3" >Approach and Goals</font></B></a></td>
<td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Method.html"><B><font face=cursive color="#FF6633" size="3">Method</font></B></a></td>
<td cellspacing="10" cellpadding="10" width="1300" width="180" bgcolor="#BAD3FF"><a href="fit_Results and Discussion.html"><B><font face=cursive color="#003366" size="3">Results and Discussion</font></B></a></td>
<td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Member.html"><B><font face=cursive color="#003366" size="3">Member</font></B></a></td>
<td cellspacing="10" cellpadding="10" width="1055" width="180" bgcolor="#BAD3FF"><a href="fit_Sponsor.html"><B><font face=cursive color="#003366" size="3">Sponsor</font></B></a></td>    </tr></table>


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<h2 p style="background:#FFFFFF; color:#003366;"><font face=cursive size="5"><B> Method </font></B></p></h2>


  <li><a href="http://openwetware.org/wiki/Biomod/2014/Fukuoka#home">Home</a>


<div style="padding: 35px;">
    <ul>


<h3><p>・Artificial experiment using fluorescent molecule<br>
      <li><a href="http://openwetware.org/wiki/Biomod/2014/Fukuoka#abs">Abstract</a></li>
<table>
      <li><a href="http://openwetware.org/wiki/Biomod/2014/Fukuoka#vid">Video</a></li>
<td><div style="width:210px;"><img alt="" src="/images//5/56/Fitfisuku.jpg" width="200" height="150" border="0"/ align="left" style="margin-right: 10px;"><p>solution of fluorescence molecular</p></div></td>
 
    </ul>
  </li>
  <li><a href="fit_Introduction.html#pro">Projects</a>
    <ul>
      <li><a href="fit_Introduction.html#back">Background & Motivation</a></li>
      <li><a href="fit_Introduction.html#goal">Project Goals</a></li>


    </ul>
  </li>
  <li><a href="fit_Approach and Goals.html#des">Design</a>
    <ul>
      <li><a href="fit_Approach and Goals.html#ear">Early Design</a></li>
      <li><a href="fit_Approach and Goals.html#fin">Final Design</a></li>
     
    </ul>
  </li>
  <li><a href="fit_Method.html#met">Method</a>
    <ul>
      <li><a href="fit_Method.html#a">Preliminary Experiment</a></li>
      <li><a href="fit_Method.html#b">Synthesis of the “Barrel” particles and the “Doll” particles</a></li>
      <li><a href="fit_Method.html#c">Combining the Doll particles with the Barrels particles</a></li>
　　　<li><a href="fit_Method.html#d">Pop-up of the doll particle</a></li>
　　　<li><a href="fit_Method.html#e">Materials</a></li>
  </ul>
  </li>
  <li><a href="fit_Results and Discussion.html#">Result and Discassions</a>
    <ul>
      <li><a href="fit_Results and Discussion.html#a">Preliminary Experiment</a></li>
      <li><a href="fit_Results and Discussion.html#b">Synthesis of the “Barrel” particles and the “Doll” particles</a></li>
      <li><a href="fit_Results and Discussion.html#c">Combining the Doll particles with the Barrels particles</a></li>
      <li><a href="fit_Results and Discussion.html#d">Pop-up of the doll particle</a></li>
　　　<li><a href="fit_Results and Discussion.html#e">Conclusion</a></li>
    </ul>
  </li>
  <li><a href="fit_Member.html#team">Team</a>
    <ul>
      <li><a href="fit_Member.html#men">Member</a></li>
      <li><a href="fit_Member.html#spo">Sponsor</a></li>
     
    </ul>
  </li>
</ul>


<td>
&nbsp; Actually, when observe it, tested it using the material which is similar that I adorned it to DNA to confirm it how much the density of fluorescence molecules can measure if there is it. The fluorescence molecules which it modified to DNA is FITC and TAMURA、Fluorescein in substitution for FITC, used Rhodamin B in substitution for TAMURA in artificial experiment.<br clear="all"></td></table>
</p><br>


<p>・Synthesis of meso porous silica particle<br>


<a name="a"></a>
<div align="left" style="margin: 1px 60px;">
<center><font size="6" color="#000022" face="Arial"><b>Preliminary Experiment<br><br>verification of FRET with the ssDNAs not modified to the silica</b></font></center>


<table>
<font size="4" color="#000022" face="Arial">
<td><div style="width:210px;"><img alt="" src="/images//7/71/Fitsilicanh.jpg" width="200" height="150" border="0"/ align="left" style="margin-right: 10px;"><p>MSN-NH2 powder</p></div></td>


<p>
&nbsp;<br>
&nbsp;We firstly confirmed that FRET is surely induced by hybridization of the sword-DNA and the barrel-DNA not yet modified on the silica. We prepared the mixed solutions of FITC/TAMRA-DNA and FITC-DNA/TAMRA-DNA. The concentration is set to 50pM. FITC-DNA/TAMRA-DNA solution was annealed: the solution was kept at 94ºC for 30 seconds and then the temperature was decreased at the rate of 6 ºC degrees per 15 minutes. Fluorescence spectra was measured on Hitachi F-2500 spectrophotometer.<br>




<td>
&nbsp; N-cetyltrimethylammonium bromide (CTAB, 1.0009 g) was first dissolved in 50 ml of pure water by heating. After cooling to room temperature, aqueous ammonia (13 ml)　and ethanol (75 ml) were added. The mixture was stirred for 15 min and TEOS (1.94 ml) added rapidly while stirring was continued. TEOS (30 μl) and APTES (30 μl) were introduced later. The mixture was allowed to stir for 2 h to give rise to white precipitates. The solid product was filtered, washed with deionized water and methanol, and dried in air. meso porous silica is synthesized Particle and pour site was determined by SEM observation.To remove the surfactant template (CTAB), the white powder was refluxed for 16 h in a solution of 1.00 ml of HCl (37%) and 80.00 ml of methanol followed by extensively washing with deionized water and methanol. Afterwards when we absorb it and filter it. After absorbing it, having filtered it, we remove water by putting meso porous silica in the vacuum state. The resulting surfactant-removed amine-functionalized MSN (MSN-NH2) was placed under high vacuum to remove the remaining solvent in the mesopores. The MSN-NH2 (50 mg) was reacted with succinic anhydride (1.00 g) in N,N-dimetylformamide solution (20 ml) under N2 gas for 8 h with continuous stirring.After a thorough water wash, the carboxylated nanoparticles (MSN-COOH) were activated using EDC (0.1471g) and sulfo-NHS (0.132g) in a MES buffer (pH 6.0) for 15 min at room temperature with continuous stirring. Twenty microliters of PBS buffer (100mM, pH7.4) was then added in the mixture, followed by the addition of TAMRA labeled DNA (5μl 100μM) at room temperature with continuous stirring for 6 h and washing in PBS buffer (0.1 M,pH 7.4) to form the resultant DNA-conjugated nanoparticles (MSN-DNA or -cDNA).
<br>
　　<br clear="all"></td></table>
</p>
</p>
　　
<p>・Characterization<br>
confocal microscope <br>
spectrophotometer


<p>
<hr>


<a name="b"></a>
<center><font size="6" color="#000022" face="Arial"><b>Synthesis of the “Barrel” particles and the “Doll” particles</b></font></center>


</p></h3>
<p>
&nbsp;  <b><1> Synthesis of mesoporous silica modified with amino group by sol-gel method.</b><br>
&nbsp; We used mesoporous silica particles to use as “Barrel” and "Doll" bodies. We used sol-gel method to synthesize the mesoporous silica modified with amino group (MPS-NH2) as shown in Scheme 1. N-cetyltrimethylammonium bromide (CTAB, 1.0009 g) was first dissolved in 50 mL of pure hot water (100 ºC). After cooling to room temperature, aqueous ammonia (28wt%, 13 mL) and ethanol (75 mL) were added. The mixture was stirred for 15 min and tetraethoxysilane (TEOS ; 1.94 ml) was added rapidly while stirring was continued. After 30 min of stirring, TEOS (0.030 mL) and aminopropyltriethoxysilane (APTES; 0.030 mL) were added. The mixture was allowed to stir for 2 h to give rise to white precipitates, followed by washing with water by using Kiriyama Rohto.<br>
<br>
<br>


<table align="left">
<td><div style="width: 330px;"><img alt="" src="/images/8/85/Fit0031.JPG" width="320" height="240" border="0" / ><p><div style="width: 330px;"><img alt="" src="/images/d/d0/Hy8.png" width="320" height="240" border="0" / ><p>  Scheme 1. Synthetic scheme for the  amino-funcionalized mesoporous silica (MPS-NH2),  carboxylated mesoporous silica (MPS-COOH), and  DNA-modified mesoporous silica.</p><div></td><tr></table>
<br>
<br>






</div>
<b><2> Modification of the mesoporous silica particle with DNA</b><br>
&nbsp; Next, we modified a carboxyl group onto the mesoporous silica with amino group. The MPS-NH2 (50 mg) was reacted with succinic anhydride (1.00 g) in N,N-dimetylformamide solution (20 mL) under N2 atmosphere for 8 h with continuous stirring in a 50 mL screw pipe. After the reaction, we washed it carefully with pure water and the carboxylated mesoporous silica (MPS- COOH) was obtained.<br>
Finally, we modified MPS-COOH with NH2-terminated ssDNAs. The MSN-COOH was dispersed in water and then activated by adding 0.1471 g of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC) and 0.1532g of N-Hydroxysuccinimide (NHS) in a MES buffer (pH 6.0) for 15 min at room temperature with continuous stirring. 0.020 mL of PBS buffer (100mM, pH 7.4) was then added in the mixture. After adding 0.005 mL of an aqueous DNA solution, the mixture was continuously stirred for 6 h at room temperature with continuous stirring. At last, we washed the sample with PBS buffer (100mM,pH7.4) three times. EDC and NHS were used to catalyze the reaction and PBS was used just for pH adjustment.<br>
<a name="c"></a>
<br>
<br>
<center><font size="6" color="#000022" face="Arial"><b> Combining the Doll particles with the Barrels particles</b></font></center>
<a name="d"></a>
<p>
&nbsp;  the Barrel particles (0.010mL) and the Doll particles (0.090mL) were mixed using a micropipette. After mixed solution was sonicated for twenty minutes<br>
<br>
<center><font size="6" color="#000022" face="Arial"><b>Pop-up of the doll particle by adding sword DNA</b></font></center>
<p>
&nbsp;  We added 0.010 mL of the sword-DNA aqueous solution to the aqueous dispersions of the barrel particles (0.010mL) followed by observation with the confocal laser scanning microscope. <br>
<br>
<a name="e"></a>
<center><font size="6" color="#000022" face="Arial"><b>Materials</b></font></center>
<p>
&nbsp;  The DNA samples shown in Table 2 were supplied from sigma-aldrich Japan. Other materials used in this project are summarized in Table 4.<br>
<br>
<center>
<table style="background-color:#ffffff;filter:alpha(opacity= 70);-moz-opacity: 0.7;opacity: 0.7;"
border="1" align="center" width="90%" height="1045" hspace="3" vspace="3" bordercolor="#666666">
<caption align="left">Table 3 .</caption>
<tr>
<th align="center" valign="middle" width="200" height="5">Structural formula</th>
<th align="center" valign="middle" width="200" height="5">Name</th>
<th align="center" valign="middle" width="200" height="5">Manufacturer</th>
<th align="center" valign="middle" width="100" height="5">Grade</th>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/2/2c/1.png.gif"/  width="160" height="100" border=""/></td>
<td align="center" valign="middle" width="200" height="130">Sulfo-NHS</td>
<td align="center" valign="middle" width="200" height="130">DOUJINDO</td>
<td align="center" valign="middle" width="100" height="130">200mg
</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/3/3b/FIT%EF%BC%92.png.gif" width="180" height="90" border=""/></td>
<td align="center" valign="middle" width="200" height="130">Tetraethyl orthosilicate<br>
(Tetraethoxysilane; TEOS)
</td>
<td align="center" valign="middle" width="200" height="130">Wako</td>
<td align="center" valign="middle" width="100" height="130">500mg</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/9/98/FIT%EF%BC%93.png.gif" width="180" height="90" border="" /></td>
<td align="center" valign="middle" width="200" height="50">3-aminopropyltriethoxysilane</td>
    <td align="center" valign="middle" width="200" height="50">Wako</td>
    <td align="center" valign="middle" width="100" height="50">25g</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/1/1f/FIT%EF%BC%94.png.png" width="230" height="120" border=""/></td>
<td align="center" valign="middle" width="200" height="130">1 - (3-dimethylaminopropyl) carbodiimide hydrochloride</td>
<td align="center" valign="middle" width="200" height="130">TGI</td>
<td align="center" valign="middle" width="100" height="130">5g</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/c/cf/FIT%E3%80%80%EF%BC%95.gif" width="150" height="100" border=""/></td>
<td align="center" valign="middle" width="200" height="130">N-N-dimethylformamide</td>
<td align="center" valign="middle" width="200" height="130">TGI</td>
<td align="center" valign="middle" width="100" height="130">200mg</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/0/0a/FIT_6.gif" width="145" height="90" border=""/></td>
<td align="center" valign="middle" width="200" height="130">Succinic anhydride</td>
<td align="center" valign="middle" width="200" height="130">Wako</td>
<td align="center" valign="middle" width="100" height="130">25g</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/7/73/FIT_7.gif" width="200" height="70" border=""/></td>
<td align="center" valign="middle" width="200" height="130">Cetyltrimethylammonium bromide</td>
<td align="center" valign="middle" width="200" height="130">Wako</td>
<td align="center" valign="middle" width="100" height="130">25g</td>
</tr>
<tr>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/c/c2/FIT_8.gif" width="180" height="90" border=""/></td>
<td align="center" valign="middle" width="200" height="130">MES
</td><td align="center" valign="middle" width="200" height="130">DOUJINDO
</td><td align="center" valign="middle" width="100" height="130">25g
</td>
</tr>
</table>


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<center><font size="6" color="#000022" face="Arial"><b>Characterization</b></font></center><br><br>


&nbsp; The synthesized particles were observed with Keyence VE7800 scanning electron microscope (SEM). We observed the synthesized particles by Nikon A1R+ confocal laser scanning microscope operated with the Ar+ laser (488 nm) and with the Galvano mirror scanner at the scanning speed of 0.125 images per second. The appratus used in this project is summarized in Table 4.
<table style="background-color:#ffffff;filter:alpha(opacity= 70);-moz-opacity: 0.7;opacity: 0.7;"  border="1" align="center" width="90%" height="440" hspace="3" vspace="3" bordercolor="#666666">
<caption align="left">Table 4 .</caption>
<tr>
<th align="center" valign="middle" width="100" height="50">Device name</th>
<th align="center" valign="middle" width="100" height="50">Manufacturer
Model number
</th>
<th align="center" valign="middle" width="200" height="50">Measurement condition </th>
<th align="center" valign="middle" width="200" height="50">Photograph</th>
        <th align="center" valign="middle" width="200" height="50">Explanations of equipment</th>
</tr>
<tr>
<td align="center" valign="middle" width="100" height="130">Spectrophotometer</td>
<td align="center" valign="middle" width="100" height="130">HITACHI F-2500</td>
<td align="center" valign="middle" width="200" height="130">WL Range:
450nm~650nm
Scanning Speed:
60nm/min
  </td>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/6/6d/FIT_9.png" width="150" height="130" border=""/>  </td>
　　　　　<td align="center" valign="middle" width="200" height="130">This device can see the concentration of the solution, the material characteristics, the structure of the molecule.  </td>
</tr>
<tr>
<td align="center" valign="middle" width="100" height="130">SEM</td>
<td align="center" valign="middle" width="100" height="130">KEYENC VE-7800</td>
<td align="center" valign="middle" width="200" height="130">WL Range:
450nm~650nm
Scanning Speed:
60nm/min
  </td>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/9/9f/FIT_10.png" width="150" height="130" border="">  </td>
　　　　　<td align="center" valign="middle" width="200" height="130">It is possible to significantly change the magnification easily. In addition, photography is possible.  </td>
</tr>
<tr>
<td align="center" valign="middle" width="100" height="130">Confocal laser scanning microscope
(CLSM)
</td>
<td align="center" valign="middle" width="100" height="130">Nikon ECLIPSE-Ti</td>
<td align="center" valign="middle" width="200" height="130">Ar-Laser (488nm)
Garvano mirror scanner
  </td>
<td align="center" valign="middle" width="200" height="130"><img alt="" src="/images/8/8e/FIT_11.png" width="150" height="130" border=""></td>
　　　　　<td align="center" valign="middle" width="200" height="130">It is possible to obtain a high resolution image in focus in the field of view for the entire sample surface with irregularities.
Three-dimensional shape measurement non-destructive, non-contact is possible
  </td>
</tr>
</table>




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  • <a href="http://openwetware.org/wiki/Biomod/2014/Fukuoka#home">Home</a>
  • <a href="fit_Introduction.html#pro">Projects</a>
    • <a href="fit_Introduction.html#back">Background & Motivation</a>
    • <a href="fit_Introduction.html#goal">Project Goals</a>
  • <a href="fit_Approach and Goals.html#des">Design</a>
    • <a href="fit_Approach and Goals.html#ear">Early Design</a>
    • <a href="fit_Approach and Goals.html#fin">Final Design</a>
  • <a href="fit_Method.html#met">Method</a>
    • <a href="fit_Method.html#a">Preliminary Experiment</a>
    • <a href="fit_Method.html#b">Synthesis of the “Barrel” particles and the “Doll” particles</a>
    • <a href="fit_Method.html#c">Combining the Doll particles with the Barrels particles</a>
      • 　　　
    • <a href="fit_Method.html#d">Pop-up of the doll particle</a>
      • 　　　
    • <a href="fit_Method.html#e">Materials</a>
  • <a href="fit_Results and Discussion.html#">Result and Discassions</a>
    • <a href="fit_Results and Discussion.html#a">Preliminary Experiment</a>
    • <a href="fit_Results and Discussion.html#b">Synthesis of the “Barrel” particles and the “Doll” particles</a>
    • <a href="fit_Results and Discussion.html#c">Combining the Doll particles with the Barrels particles</a>
    • <a href="fit_Results and Discussion.html#d">Pop-up of the doll particle</a>
      • 　　　
    • <a href="fit_Results and Discussion.html#e">Conclusion</a>
  • <a href="fit_Member.html#team">Team</a>
    • <a href="fit_Member.html#men">Member</a>
    • <a href="fit_Member.html#spo">Sponsor</a>


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Preliminary Experiment

verification of FRET with the ssDNAs not modified to the silica

 
 We firstly confirmed that FRET is surely induced by hybridization of the sword-DNA and the barrel-DNA not yet modified on the silica. We prepared the mixed solutions of FITC/TAMRA-DNA and FITC-DNA/TAMRA-DNA. The concentration is set to 50pM. FITC-DNA/TAMRA-DNA solution was annealed: the solution was kept at 94ºC for 30 seconds and then the temperature was decreased at the rate of 6 ºC degrees per 15 minutes. Fluorescence spectra was measured on Hitachi F-2500 spectrophotometer.

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Synthesis of the “Barrel” particles and the “Doll” particles

  <1> Synthesis of mesoporous silica modified with amino group by sol-gel method.
  We used mesoporous silica particles to use as “Barrel” and "Doll" bodies. We used sol-gel method to synthesize the mesoporous silica modified with amino group (MPS-NH2) as shown in Scheme 1. N-cetyltrimethylammonium bromide (CTAB, 1.0009 g) was first dissolved in 50 mL of pure hot water (100 ºC). After cooling to room temperature, aqueous ammonia (28wt%, 13 mL) and ethanol (75 mL) were added. The mixture was stirred for 15 min and tetraethoxysilane (TEOS ; 1.94 ml) was added rapidly while stirring was continued. After 30 min of stirring, TEOS (0.030 mL) and aminopropyltriethoxysilane (APTES; 0.030 mL) were added. The mixture was allowed to stir for 2 h to give rise to white precipitates, followed by washing with water by using Kiriyama Rohto.


<img alt="" src="/images/8/85/Fit0031.JPG" width="320" height="240" border="0" / >

<img alt="" src="/images/d/d0/Hy8.png" width="320" height="240" border="0" / >

Scheme 1. Synthetic scheme for the amino-funcionalized mesoporous silica (MPS-NH2), carboxylated mesoporous silica (MPS-COOH), and DNA-modified mesoporous silica.




<2> Modification of the mesoporous silica particle with DNA
  Next, we modified a carboxyl group onto the mesoporous silica with amino group. The MPS-NH2 (50 mg) was reacted with succinic anhydride (1.00 g) in N,N-dimetylformamide solution (20 mL) under N2 atmosphere for 8 h with continuous stirring in a 50 mL screw pipe. After the reaction, we washed it carefully with pure water and the carboxylated mesoporous silica (MPS- COOH) was obtained.
Finally, we modified MPS-COOH with NH2-terminated ssDNAs. The MSN-COOH was dispersed in water and then activated by adding 0.1471 g of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC) and 0.1532g of N-Hydroxysuccinimide (NHS) in a MES buffer (pH 6.0) for 15 min at room temperature with continuous stirring. 0.020 mL of PBS buffer (100mM, pH 7.4) was then added in the mixture. After adding 0.005 mL of an aqueous DNA solution, the mixture was continuously stirred for 6 h at room temperature with continuous stirring. At last, we washed the sample with PBS buffer (100mM,pH7.4) three times. EDC and NHS were used to catalyze the reaction and PBS was used just for pH adjustment.
<a name="c"></a>

Combining the Doll particles with the Barrels particles

<a name="d"></a>

  the Barrel particles (0.010mL) and the Doll particles (0.090mL) were mixed using a micropipette. After mixed solution was sonicated for twenty minutes

Pop-up of the doll particle by adding sword DNA

  We added 0.010 mL of the sword-DNA aqueous solution to the aqueous dispersions of the barrel particles (0.010mL) followed by observation with the confocal laser scanning microscope.

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Materials

  The DNA samples shown in Table 2 were supplied from sigma-aldrich Japan. Other materials used in this project are summarized in Table 4.

Table 3 .
Structural formula Name Manufacturer Grade
<img alt="" src="/images/2/2c/1.png.gif"/ width="160" height="100" border=""/> Sulfo-NHS DOUJINDO 200mg
<img alt="" src="/images/3/3b/FIT%EF%BC%92.png.gif" width="180" height="90" border=""/> Tetraethyl orthosilicate

(Tetraethoxysilane; TEOS)

Wako 500mg
<img alt="" src="/images/9/98/FIT%EF%BC%93.png.gif" width="180" height="90" border="" /> 3-aminopropyltriethoxysilane Wako 25g
<img alt="" src="/images/1/1f/FIT%EF%BC%94.png.png" width="230" height="120" border=""/> 1 - (3-dimethylaminopropyl) carbodiimide hydrochloride TGI 5g
<img alt="" src="/images/c/cf/FIT%E3%80%80%EF%BC%95.gif" width="150" height="100" border=""/> N-N-dimethylformamide TGI 200mg
<img alt="" src="/images/0/0a/FIT_6.gif" width="145" height="90" border=""/> Succinic anhydride Wako 25g
<img alt="" src="/images/7/73/FIT_7.gif" width="200" height="70" border=""/> Cetyltrimethylammonium bromide Wako 25g
<img alt="" src="/images/c/c2/FIT_8.gif" width="180" height="90" border=""/> MES DOUJINDO 25g




]



Characterization


  The synthesized particles were observed with Keyence VE7800 scanning electron microscope (SEM). We observed the synthesized particles by Nikon A1R+ confocal laser scanning microscope operated with the Ar+ laser (488 nm) and with the Galvano mirror scanner at the scanning speed of 0.125 images per second. The appratus used in this project is summarized in Table 4.

　　　　　 　　　　　 　　　　　
Table 4 .
Device name Manufacturer

Model number

Measurement condition Photograph Explanations of equipment
Spectrophotometer HITACHI F-2500 WL Range:

450nm~650nm Scanning Speed: 60nm/min

<img alt="" src="/images/6/6d/FIT_9.png" width="150" height="130" border=""/> This device can see the concentration of the solution, the material characteristics, the structure of the molecule.
SEM KEYENC VE-7800 WL Range:

450nm~650nm Scanning Speed: 60nm/min

<img alt="" src="/images/9/9f/FIT_10.png" width="150" height="130" border=""> It is possible to significantly change the magnification easily. In addition, photography is possible.
Confocal laser scanning microscope

(CLSM)

Nikon ECLIPSE-Ti Ar-Laser (488nm)

Garvano mirror scanner

<img alt="" src="/images/8/8e/FIT_11.png" width="150" height="130" border="">It is possible to obtain a high resolution image in focus in the field of view for the entire sample surface with irregularities.

Three-dimensional shape measurement non-destructive, non-contact is possible






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