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 |
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##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
Now that I have the images before the wash steps, I continued with the normal wash steps for all the probes.
Hybridization
- Remove coverslip carefully and wash in pre-warmed 2X SSCT at 60C for 15 minutes
- Incubate in 2X SSCT at room temperature for 10 minutes
- Incubate in 0.2X SSC at room temperature for 10 minutes
- Dry with kimwipes; do not let the surface containing the cells touch the kimwipes
- Add 15 uL DRAQ5 (diluted 1:1000) to center of dish
- Cover with 22x22 #1.5 coverslip; seal with nail polish
- Allow at least 30 minutes for nail polish to dry
- Slides are quite stable for several months at 4C
Microscopy
Sample D
- 20140807 Cot1488.lif Cot1448 1 ch00.jpg
Cells stained with Cot1, Alexa 488 (green), field 1
- 20140807 Cot1488.lif Cot1488 2 ch00.jpg
Cells stained with Cot1, Alexa 488 (green), field 2
- 20140807 Cot1488.lif Cot1488 3 ch00.jpg
Cells stained with Cot1, Alexa 488 (green), field 2
Sample A
- 20140807 DMD488.lif DMD488 1 ch00.jpg
Cells stained with DMD, Alexa 488 (green), field 1
- 20140807 DMD488.lif DMD488 2 ch00.jpg
Cells stained with DMD, Alexa 488 (green), field 2
Sample B
- 20140807 DMD546 DMD546 1 ch00.jpg
Cells stained with DMD, Alexa 546 (red), field 1
- 20140807 DMD546 DMD546 2 ch00.jpg
Cells stained with DMD, Alexa 546 (red), field 2
Sample C
- 20140807 DMD488 Dilute.lif DMD488 D1 ch00.jpg
Cells stained with DMD, Alexa 488 (green), field 1
- 20140807 DMD488 Dilute.lif DMD488 D2 ch00.jpg
Cells stained with DMD, Alexa 488 (green), field 2