Hosuk:LabNotes/2014-8-19: Difference between revisions
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*Step0 : 1st Rolony --> ch00 : Cy3, ch01 : BF | *Step# after confocal imaging | ||
*Step1~7 : ch00 : 488, ch01 : Cy3, ch02 : Cy5, ch03 : BF | **Step0 : 1st Rolony --> ch00 : Cy3, ch01 : BF | ||
**Step1~7 : ch00 : 488, ch01 : Cy3, ch02 : Cy5, ch03 : BF | |||
*Maximum Intensity Projection | *Maximum Intensity Projection | ||
**Re-format the 'Step#' --> Step1~21 | |||
**Step1 --> Step1:488, Step2:Cy3, Step3:Cy5 | |||
**Step2 --> Step4:488, Step5:Cy3, Step6:Cy5 | |||
**...Step7 --> Step19:488, Step20:Cy3, Step21:Cy5 | |||
*Image aligning | *Image aligning | ||
*#Shift, Crop | *#Shift, Crop |
Revision as of 21:04, 22 August 2014
Decoding with Agi26k0gap Padlock Probe
- Image resolution : 20x obj, 4096 x 4086, z step = 1.04um
- Since how many features would be detected or how much dense rolonies in this sample, so image in wide field of view
- eventually 63x obj.
Analysis
Arrange data
- File name (decided at imaging) : [Sample]_[Resolution]_[Step#]_[Pos#]_z##_ch##.tif
- ex) PGP1F : PGP1F_Agi26k0gap_Decode_P20_S2_2014-08-09_20x_4k_Step1_Pos2_z01_ch01.tif
- ex) Tissue section : HBTissue_2014-08-12_RCAagain_20x_4k_Pos1_z00_ch00.tif
- Step# after confocal imaging
- Step0 : 1st Rolony --> ch00 : Cy3, ch01 : BF
- Step1~7 : ch00 : 488, ch01 : Cy3, ch02 : Cy5, ch03 : BF
- Maximum Intensity Projection
- Re-format the 'Step#' --> Step1~21
- Step1 --> Step1:488, Step2:Cy3, Step3:Cy5
- Step2 --> Step4:488, Step5:Cy3, Step6:Cy5
- ...Step7 --> Step19:488, Step20:Cy3, Step21:Cy5
- Image aligning
- Shift, Crop
- Shift
- BF
Result
- PGP1F_P20_S2_2014-08-09 --> , HBTissue_2014-08-12_S1