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Daniel:Notebook/HiResChrPaint/2014-8-7
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=No Wash FISH= [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] Now that I have the [[Daniel:Notebook/HiResChrPaint/2014-8-6|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=== <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> [[Category:HRCP]] [[Category:Hybridization]] [[Category:Microscopy]] [[Category:20140805]]
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