Daniel:Notebook/HiResChrPaint/2014-8-7: Difference between revisions

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File:20140807_DMD488_Dilute.lif_DMD488_D2_ch00.jpg|Cells stained with DMD, Alexa 488 (green), field 2
File:20140807_DMD488_Dilute.lif_DMD488_D2_ch00.jpg|Cells stained with DMD, Alexa 488 (green), field 2
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[[Category:HRCP]] [[Category:Hybridization]] [[Category:Microscopy]] [[Category:20140805]]

Latest revision as of 18:56, 12 August 2014

No Wash FISH[edit]

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Now that I have the images before the wash steps, I continued with the normal wash steps for all the probes.

Hybridization[edit]

  1. Remove coverslip carefully and wash in pre-warmed 2X SSCT at 60C for 15 minutes
  2. Incubate in 2X SSCT at room temperature for 10 minutes
  3. Incubate in 0.2X SSC at room temperature for 10 minutes
  4. Dry with kimwipes; do not let the surface containing the cells touch the kimwipes
  5. Add 15 uL DRAQ5 (diluted 1:1000) to center of dish
  6. Cover with 22x22 #1.5 coverslip; seal with nail polish
    1. Allow at least 30 minutes for nail polish to dry
    2. Slides are quite stable for several months at 4C

Microscopy[edit]

Sample D[edit]

Sample A[edit]

Sample B[edit]

Sample C[edit]