Daniel:Notebook/HiResChrPaint/2014-8-12
FISH with DAPI (Started 08/10/2014)
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 C
- 20140812 Cot1488.lif Field1 ch00.jpg
NCI h1975 cells stained with Cot1-488 (green); field 1
- 20140812 Cot1488.lif Field2 ch00.jpg
NCI h1975 cells stained with Cot1-488 (green); field 2
Sample A
- 20140812 DMD488.lif Field1 ch00.jpg
NCI h1975 cells stained with DMD-488 (green); field 1
- 20140812 DMD488.lif Field2 ch00.jpg
NCI h1975 cells stained with DMD-488 (green); field 2
Sample B
- 20140812 DMD546.lif Field1 ch00.jpg
NCI h1975 cells stained with DMD-546 (red); field 1
- 20140812 DMD546.lif Field2 ch00.jpg
NCI h1975 cells stained with DMD-546 (red); field 2
Sample D <gallery> File:20140812_Cot1488DMD546.lif_Field1_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 1 (3.55X) File:20140812_Cot1488DMD546.lif_Field2_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 2 (3.55X) File:20140812_Cot1488DMD546.lif_Field2_1X_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 2 (1X) File:20140812_Cot1488DMD546.lif_Field3_ch00.jpg|NCI h1975 cells stained with DMD-546 (red) and Cot1-488 (green); field 3 (5X) <gallery>