User:Andy Maloney/Notebook/Lab Notebook of Andy Maloney/2009/11/25/Sucrose: Difference between revisions
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* I made my [http://www.openwetware.org/wiki/User:Andy_Maloney/Kinesin_%26_Microtubule_Page#Flow_cell flow cells]. | * I made my [http://www.openwetware.org/wiki/User:Andy_Maloney/Kinesin_%26_Microtubule_Page#Flow_cell flow cells]. | ||
==Motility== | ==Motility== | ||
So I did change some things up from my heavy water experiments. Instead of doing the sucrose dilutions sequentially in time, I prepared all of them at the same time. I did however, not flow in the microtubules until after I took data for | So I did change some things up from my heavy water experiments. Instead of doing the sucrose dilutions sequentially in time, I prepared all of them at the same time. I did however, not flow in the microtubules until after I took data for each slide. This may have caused me some issues with the 750 mM sucrose guy. I'll have to redo this to make sure. Suffice it to say though, the 250 mM sucrose guy does look like the MTs are moving slower and that's all I was hoping to see today. I will have to redo it anyway in a cleaner fashion to make sure I didn't do something silly. | ||
All movies are 2 minutes long for one field of view (I took 3 FOVs). The playback rate is 5 frames/s and the capture rate was 5 Hz so the movies are not sped up. The images have an exposure of 100 ms and a gain of 150. | |||
====250 mM Sucrose==== | ====250 mM Sucrose==== | ||
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</html> | </html> | ||
==Notes== | ==Notes== | ||
These guys definitely look like they are running slower than the motility assays I have done without osmolytes in them. Of course, I cannot determine how much slower until I analyze the movies made. | These guys definitely look like they are running slower than the motility assays I have done without osmolytes in them. Of course, I cannot determine how much slower until I analyze the movies made. | ||
Revision as of 15:26, 25 November 2009
Setup
- I polymerized microtubules.
- I put 2 M sucrose in PEM.
- I prepared from the 2 M sucrose solution, 3 dilutions of 0.25 M, 0.50 M, and 0.75 M sucrose in the motility solutions.
- I made my flow cells.
Motility
So I did change some things up from my heavy water experiments. Instead of doing the sucrose dilutions sequentially in time, I prepared all of them at the same time. I did however, not flow in the microtubules until after I took data for each slide. This may have caused me some issues with the 750 mM sucrose guy. I'll have to redo this to make sure. Suffice it to say though, the 250 mM sucrose guy does look like the MTs are moving slower and that's all I was hoping to see today. I will have to redo it anyway in a cleaner fashion to make sure I didn't do something silly.
All movies are 2 minutes long for one field of view (I took 3 FOVs). The playback rate is 5 frames/s and the capture rate was 5 Hz so the movies are not sped up. The images have an exposure of 100 ms and a gain of 150.
250 mM Sucrose
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500 mM Sucrose
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750 mM Sucrose
<html> <object width="425" height="344"><param name="movie" value="http://www.youtube.com/v/5aRu60dVlrk&hl=en_US&fs=1&"></param><param name="allowFullScreen" value="true"></param><param name="allowscriptaccess" value="always"></param><embed src="http://www.youtube.com/v/5aRu60dVlrk&hl=en_US&fs=1&" type="application/x-shockwave-flash" allowscriptaccess="always" allowfullscreen="true" width="425" height="344"></embed></object> </html>
Notes
These guys definitely look like they are running slower than the motility assays I have done without osmolytes in them. Of course, I cannot determine how much slower until I analyze the movies made.
