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

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>Djacobse
(Created page with "=No Wash FISH= Back to Calendar Now that I have the images before the wash steps, I continued wi...")
 
>Djacobse
Line 15: Line 15:
##Allow at least 30 minutes for nail polish to dry
##Allow at least 30 minutes for nail polish to dry
##Slides are quite stable for several months at 4C
##Slides are quite stable for several months at 4C
==Microscopy==
===Sample D===
<gallery>
File:20140807_Cot1488.lif_Cot1448_1_ch00.jpg|Cells stained with Cot1, Alexa 488 (green), field 1
File:20140807_Cot1488.lif_Cot1488_2_ch00.jpg|Cells stained with Cot1, Alexa 488 (green), field 2
File:20140807_Cot1488.lif_Cot1488_3_ch00.jpg|Cells stained with Cot1, Alexa 488 (green), field 2
</gallery>
===Sample A===
<gallery>
File:20140807_DMD488.lif_DMD488_1_ch00.jpg|Cells stained with DMD, Alexa 488 (green), field 1
File:20140807_DMD488.lif_DMD488_2_ch00.jpg|Cells stained with DMD, Alexa 488 (green), field 2
</gallery>
===Sample B===
<gallery>
File:20140807_DMD546_DMD546_1_ch00.jpg|Cells stained with DMD, Alexa 546 (red), field 1
File:20140807_DMD546_DMD546_2_ch00.jpg|Cells stained with DMD, Alexa 546 (red), field 2
</gallery>
===Sample C===
<gallery>
File:20140807_DMD488_Dilute.lif_DMD488_D1_ch00.jpg|Cells stained with DMD, Alexa 488 (green), field 1
File:20140807_DMD488_Dilute.lif_DMD488_D2_ch00.jpg|Cells stained with DMD, Alexa 488 (green), field 2
</gallery>

Revision as of 20:57, 7 August 2014

No Wash FISH

Back to Calendar

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

Hybridization

  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

Sample D

Sample A

Sample B

Sample C