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 No edit summary |
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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 | ||
Line 15: | Line 18: | ||
**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 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]] | |||
* | *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
- epi FL pictures (Zeiss, 20x NA=0.75, 400um x 400um, single z plane)
- File:S1 S2 MatTekDish epiFL 2013-10-22.png
- Confocal pictures
- location 1 : 1st Rolonies, ACTB
- location 2 : 1st Rolonies, ACTB
- Image processing and counting will be done with using PISA...