Biomod/2011/TUM/TNT/LabbookA/Preparing samples for transmission electron microscope: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 210: Line 210:
<br>
<br>
[http://openwetware.org/wiki/Tnt_-_Labbook/2011/09/16#prepared_grids_for_TEM 16. September] <br>
[http://openwetware.org/wiki/Tnt_-_Labbook/2011/09/16#prepared_grids_for_TEM 16. September] <br>
B2: BM2 control without DNA-binders
B2: BM2 control without DNA-binders <br>
B3: BM2 with 10µM spermine <br>
B3: BM2 with 10µM spermine <br>
B4: BM2 with 30µM spermine <br>
B4: BM2 with 30µM spermine <br>
Line 218: Line 218:
C4: BM2 with 250µM ethidium bromide <br>
C4: BM2 with 250µM ethidium bromide <br>
<br>
<br>
28. September <br>
[http://openwetware.org/wiki/Tnt_-_Labbook/2011/09/28#prepared_grids_for_TEM 28. September] <br>
C5: BM2 control without DNA-binders <br>
D1: BM2 with 6µM EtBr <br>
D2: BM2 with 12µM EtBr <br>
D3: BM2 with 2.4µM spermine <br>
D4: BM2 with 4.8µM spermine <br>
E1: BM2 with 0.9nM DAPI <br>
E2: BM2 with 1.8nM DAPI <br>
<br>

Revision as of 03:26, 28 September 2011

<html> 

  <style>
  1. column-one { display:none; width:0px;}

.container{background-color: #f5f5f5; margin-top:50px}

  1. content { width: 0px; margin: 0 auto auto 0; padding: 1em 1em 1em 1em; align: center;}
  1. column-content {width: 0px; float: left; margin: 0 0 0 0;padding: 0;}

.firstHeading {display:none; width:0px;}

  1. globalWrapper{ width:1280px;}
  1. column-one {display:none; width:0px;background-color: #ffffff;}
  1. content{ margin: 0 0 0 0; padding: 0em 1em 1em 1em; position: center; width: 800px;background-color: #f5f5f5; }

.container{ width: 800px; margin: auto; background-color: #ffffff; text-align:justify; font-family: helvetica, arial, sans-serif; color:#555555; margin-top:25px; }

  1. bodyContent{ width: 1280px; align: center; background-color: #ffffff;}
  1. column-content{width: 1280px;background-color: #ffffff;}

.firstHeading { display:none;width:0px;background-color: #ffffff;}

  1. header{position: center; width: 800px;background-color: #ffffff;}

<style type="text/css"> body { 

   font-family: helvetica, arial, sans-serif;
   color: black;

background:#f5f5f5; 

 }

ul#topnav {

margin: 0 0 0 0 ; padding: 0 0 0 0; float: left; width: 800px; list-style: none; position: relative; 

       font-family: helvetica, arial, sans-serif;

font-size: 1em; background: #f5f5f5; 

       height: 5px;

} ul#topnav li { float: left; margin: 0; padding: 0; border-right: 6px solid #ffffff; /*--Divider for each parent level links--*/ } ul#topnav li a { padding: 10px 15px; display: block; color: #333333; text-decoration: none; } ul#topnav li:hover { background: #cccccc;} /*--Notice the hover color is on the list item itself, not on the link. This is so it can stay highlighted even when hovering over the subnav--*/

ul#topnav li span { float: left; padding: 15px 0; position: absolute; left: 0; top:30px; display: none; /*--Hide by default--*/ width: 800px; background: #cccccc; color: #333333; /*--Bottom right rounded corner--*/ -moz-border-radius-bottomright: 5px; -khtml-border-radius-bottomright: 5px; -webkit-border-bottom-right-radius: 5px; /*--Bottom left rounded corner--*/ -moz-border-radius-bottomleft: 5px; -khtml-border-radius-bottomleft: 5px; -webkit-border-bottom-left-radius: 5px; } ul#topnav li:hover span { display: block; } /*--Show subnav on hover--*/ ul#topnav li span a { display: inline; } /*--Since we declared a link style on the parent list link, we will correct it back to its original state--*/ ul#topnav li span a:hover {text-decoration: underline;}


  1. abstractandlinks{

width:800px; background-color:#f5f5f5; text-align:justify;font-family: helvetica, arial, sans-serif; color:#555555; margin-top:60px; }

  1. abstract{

font-family: helvetica, arial, sans-serif; width: 490px; margin-top: 20px; padding:0px 5px; float: left;background-color:#f5f5f5; height: 410px; }

  1. links{

font-family: helvetica, arial, sans-serif; width: 290px; margin-top: 20px; padding:0px 5px; float: left; background:#f5f5f5;height: 410px; }

  1. tour{

font-family: helvetica, arial, sans-serif; width: 790px; padding:0px 5px; margin-top: 1em, margin-bottom: 1em; float: left;background:#f5f5f5; font-size: 2em; }

  1. struktur{

font-family: helvetica, arial, sans-serif; width: 790px; padding:0px 5px; float: left;background:#f5f5f5; }

</style>

<head> <title> 

       TUM NanU - Labbook

</title> </head> <body> <body background-color="#F0F0F0"> <div id="contentSub"> <span class="subpages"> < <a href="/wiki/Tnt_-_Labbook" title="Tnt - Labbook">TUM-NanU - Labbook</a> 

|

<a href="/wiki/Tnt_-_Labbook/2011" title="Tnt - Labbook/2011">2011</a> 

|

<a href="/wiki/Tnt_-_Labbook/2011/09" title="Tnt - Labbook/2011/09">09</a> </span> </div> <!-- Header--> <div class="container"> 

   <div id="header"> <img src="http://openwetware.org/images/5/5c/Header2.png" /> </div>

<!--Menue--> <ul id="topnav"> 

   <li><a href="http://openwetware.org/wiki/Biomod/2011/TUM/TNT/Home">Home</a></li>
   <li>
       <a href="#">Project</a>
       <!--Subnav Starts Here-->
       <span>
           <a href="#">The Idea</a> |
           <a href="#">The Plan</a> |
           <a href="#">The Video</a>
       </span>
       <!--Subnav Ends Here-->
   </li>
   <li><a href="#">Results</a>

<span> 

           <a href="#">Structure</a> 
           <a href="#">FRET</a> 
           <a href="#">....</a>
       </span>
       <!--Subnav Ends Here-->

</li> 

   <li><a href="http://openwetware.org/wiki/Biomod/2011/TUM/TNT">Testpage</a></li>
   <li><a href="http://openwetware.org/wiki/Biomod/2011/TUM/TNT/Team">Team</a></li>
   <li><a href="http://openwetware.org/wiki/Tnt_-_Labbook">Labbook</a><!--Subnav Starts Here-->
       <span>
           <a href="#">Week I</a> 
           <a href="#">Week II</a> 
           <a href="#">Week III</a>
       </span>
       <!--Subnav Ends Here-->
   </li>
   <li><a href="#">Outlook</a></li>
   <li><a href="#">Extras</a><span>
           <a href="#">How to do a stop-motion-video</a> 
           <a href="#">How to make ...</a> 
           <a href="http://openwetware.org/index.php?title=Tnt_-_Labbook&action=submit">Edit</a>
       </span>

</li> </ul> <p> <p> </body>


</html>




preparing samples for transmission electron microscope

glow-discharge grids at 30mA for 45s
add 1µl 5M NaOH to 200µl 2% uranyl acetate staining solution
hide solution from light
vortex 1minute
centrifuge for 2.5 minutes at maximum speed
apply 5µl sample on grid
incubate for 3 minutes
remove liquid from grid without touching its sourface
lay grid on 20µl staining solution, then remove stain immediatly from grid
lay grid on another drop of 20µl staining solution
incubate for 40s seconds
remove liquid from grid
let grid dry for at least 15 minutes

grid positions


BIOMOD grid box

30. August
A2: 2DH3ML_BM1
A3: 2DH3ML_BM2
A4: 5DH3ML_BM1
A5: 5DH3ML_BM2

16. September
B2: BM2 control without DNA-binders
B3: BM2 with 10µM spermine
B4: BM2 with 30µM spermine
B5: BM2 with 100µM spermine
C2: BM2 with 25µM ethidium bromide
C3: BM2 with 75µM ethidium bromide
C4: BM2 with 250µM ethidium bromide

28. September
C5: BM2 control without DNA-binders
D1: BM2 with 6µM EtBr
D2: BM2 with 12µM EtBr
D3: BM2 with 2.4µM spermine
D4: BM2 with 4.8µM spermine
E1: BM2 with 0.9nM DAPI
E2: BM2 with 1.8nM DAPI

Retrieved from "https://openwetware.org/mediawiki/index.php?title=Biomod/2011/TUM/TNT/LabbookA/Preparing_samples_for_transmission_electron_microscope&oldid=541993"