Hosuk:LabNotes/2013-10-22: Difference between revisions

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>Hosuki78
(Created page with "===Rolonies with fresh cells - MatTek dish=== *Produce rolonies in MatTek dishes which were fixed at 10/17, the same cells to 96 well plate. ====Process==== *2 dishes *RT :...")
 
>Hosuki78
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*[[Hosuk:Lab_Notes|LabNote]]
===Rolonies with fresh cells - MatTek dish===
===Rolonies with fresh cells - MatTek dish===


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====Process====
====Process====
*2 dishes
*2 dishes
*RT : 8:30 pm
*RT : 10/20 8:30 pm
*CircLigase : 10/21 12:00 pm
*CircLigase : 10/21 12:00 pm
*End RCA : 10/22 13:00pm
*End RCA : 10/22 13:00pm
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**Of course, the number and density of rolonies are still not as many as the sample 06/23...
**Of course, the number and density of rolonies are still not as many as the sample 06/23...


*Doing Confocal
*Doing 1 location as well as tile scanning
**Doing 1 location as well as tile scanning
**Now Confocal is doing tile scanning...
** Tile : 7 x 6 (1 tile field of view : 581.25um x 581.25um),
** Total Z scan :35.81um, step : 1.23um, Total 30 stacks
** 3 hr. 39min
 
 
*epi FL pictures (Zeiss, 20x NA=0.75, 400um x 400um, single z plane)
*[[File:S1_S2_MatTekDish_epiFL_2013-10-22.png|800px]]




*Some of pictures...
*Confocal pictures
**location 1 : [[Media:S2_20131022_20x_2kx2k_pos1_Zstack-1.2um_MaxProjection_Cy5.tif|1st Rolonies]], [[Media:S2_20131022_20x_2kx2k_pos1_Zstack-1.2um_MaxProjection_ATTO488.tif|ACTB]]
**location 1 : [[Media:S2_20131022_20x_2kx2k_pos1_Zstack-1.2um_MaxProjection_Cy5.tif|1st Rolonies]], [[Media:S2_20131022_20x_2kx2k_pos1_Zstack-1.2um_MaxProjection_ATTO488.tif|ACTB]]
**location 2 : [[Media:S2_20131022_20x_2kx2k_pos2_Zstack-1.2um_MaxProjection_Cy5.tif|1st Rolonies]], [[Media:S2_20131022_20x_2kx2k_pos2_Zstack-1.2um_MaxProjection_ATTO488.tif|ACTB]]
**location 2 : [[Media:S2_20131022_20x_2kx2k_pos2_Zstack-1.2um_MaxProjection_Cy5.tif|1st Rolonies]], [[Media:S2_20131022_20x_2kx2k_pos2_Zstack-1.2um_MaxProjection_ATTO488.tif|ACTB]]
*[[File:S1_S2_MatTekDish_Confocal_2013-10-22.png|900px]]
*Image processing and counting will be done with using PISA...

Latest revision as of 17:30, 24 October 2013


Rolonies with fresh cells - MatTek dish[edit]

  • Produce rolonies in MatTek dishes which were fixed at 10/17, the same cells to 96 well plate.

Process[edit]

  • 2 dishes
  • RT : 10/20 8:30 pm
  • CircLigase : 10/21 12:00 pm
  • End RCA : 10/22 13:00pm
  • Imaging


Result[edit]

  • Good, many rolonies, and more ACTB than before.
    • Of course, the number and density of rolonies are still not as many as the sample 06/23...
  • Doing 1 location as well as tile scanning
    • Now Confocal is doing tile scanning...
    • Tile : 7 x 6 (1 tile field of view : 581.25um x 581.25um),
    • Total Z scan :35.81um, step : 1.23um, Total 30 stacks
    • 3 hr. 39min





  • Image processing and counting will be done with using PISA...