Hosuk:LabNotes/2015-3-26

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Imaging with Structural Illumination microscopy[edit]

  • New microscope in Yun’s lab : Zeiss, Apotome2 structural illumination microscopy
  • Image with 3 samples
    • PGP1F_S1_1st Rolony_2015-01-18
    • BA8_S1_1st Rolony_2015-02-27
    • BA8_S1_V4_2015-02-14


  • SIL imaging condition
    • 20x obj., Image size : 2752 x 2208 pixel, 0.23 um /pixel
    • --> 632.96 x 507.84 um
  • Brain tissue section has a strong auto-fluoresence!, so we need image with 488 channel as a auto fluorescent signal.
    • Auto-fluorescent signal image(488 channel) was subtract from BA8, 2015-02-14 Image, but not from BA8, 2015-02-27 Image (we didn’t have enough time to re-image). So BA8, 2015-02-27 image potentially have auto-fluorescent image!



Images[edit]

Sample:PGP1F_S1_2015-01-18[edit]

File:Imging 2015-03-26 PGP1F S1 2015-01-18 Zeiss SIL 20x.png


File:Imging 2015-03-26 PGP1F S1 2015-01-18 Zeiss SIL 20x Detail1.png


Sample:BA8_1stRolony_S1_2015-02-27[edit]

File:Imging 2015-03-26 BA8 1stRolony S1 2015-02-27 Zeiss SIL 20x.png


File:Imging 2015-03-26 BA8 1stRolony S1 2015-02-27 Zeiss SIL 20x Detail1.png


Sample:BA8_CA12kNov_V4_S1_2015-02-14[edit]

File:Imging 2015-03-26 BA8 CA12kNov V4 S1 2015-02-14 Zeiss SIL 20x.png


File:Imging 2015-03-26 BA8 CA12kNov V4 S1 2015-02-14 Zeiss SIL 20x Detail1.png



Comparison[edit]

  • Imaging time (20x obj, 10 stacks, 3 colors)
    • Confocal 2048 x 2048 px : ~ 40 sec
    • Confocal 4096 x 4096 px : ~ 2min 40 sec
    • SIL 2752 x 2208 px : ~ 30 sec


  • Area of a field of view (20x obj, 10 stacks, 3 colors)
    • Confocal 2048 x 2048 px : 581 um x 581 um
    • Confocal 4096 x 4096 px : 581 um x 581 um
    • SIL 2752 x 2208 px : 632.96 x 507.84 um


  • Resolution quality :
    • Confocal 2048 x 2048 < SIL 20x < Confocal 4096 x 4096


File:Comparison Leica Confocal 20x 4k 2k Zeiss SIL 20x 2.png