Editing
Daniel:Notebook/HiResChrPaint/2014-8-23
(section)
Jump to navigation
Jump to search
Warning:
You are not logged in. Your IP address will be publicly visible if you make any edits. If you
log in
or
create an account
, your edits will be attributed to your username, along with other benefits.
Anti-spam check. Do
not
fill this in!
=Padlock Probes ([[Daniel:Notebook/HiResChrPaint/2014-8-13|Started 8/13/2014]])= [[ Daniel:Notebook/HiResChrPaint|Back to Calendar]] ==Hybridization ([[Daniel:Notebook/HiResChrPaint/2014-8-21|From 8/21/2014]])== #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 20 uL DAPI + Slow Fade Gold 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 A=== Sample A is stained with PPRL-488 (green) and Cot1-546 (red). The padlock probes should produce strong green signal in many cells, but there is only weak signal in a very small number of cells. <gallery> File:20140823_PPRL488_Cot1546.lif_Field1_ch00.jpg|Cells stained with PPRL-488 (green) and Cot1-546 (red); field 1 File:20140823_PPRL488_Cot1546.lif_Field1_Zoom_ch00.jpg|Cells stained with PPRL-488 (green) and Cot1-546 (red); field 1 zoom File:20140823_PPRL488_Cot1546.lif_Field2_ch00.jpg|Cells stained with PPRL-488 (green) and Cot1-546 (red); field 2 File:20140823_PPRL488_Cot1546.lif_Field2_Zoom_ch00.jpg|Cells stained with PPRL-488 (green) and Cot1-546 (red); field 2 zoom File:20140823_PPRL488_Cot1546.lif_Field3_ch00.jpg|Cells stained with PPRL-488 (green) and Cot1-546 (red); field 3 </gallery> ===Sample B=== Sample B is stained with Cot1-488 (green) and DMD-546 (red). The signal seems to be present in several cells, though it is difficult to determine, given the small scale of the signal (the probes target only a few kb). Zooming in to get only one or two cells helps detect the signal better. However, there does appear to be clear signal above the noise in #1 and #3. #2 is meant to show what most cells look like, containing strong Cot1 signal with very no signal from the DMD. <gallery> File:20140823_Cot1488_DMD546.lif_Field1_ch00.jpg|Cells stained with Cot1-488 (green) and DMD-546 (red); field 1 File:20140823_Cot1488_DMD546.lif_Field2_ch00.jpg|Cells stained with Cot1-488 (green) and DMD-546 (red); field 2 File:20140823_Cot1488_DMD546.lif_Field3_ch00.jpg|Cells stained with Cot1-488 (green) and DMD-546 (red); field 3 </gallery> [[Category:HRCP]] [[Category:Hybridization]] [[Category:Microscopy]] [[Category:20140813]]
Summary:
Please note that all contributions to ZhangLabWiki may be edited, altered, or removed by other contributors. If you do not want your writing to be edited mercilessly, then do not submit it here.
You are also promising us that you wrote this yourself, or copied it from a public domain or similar free resource (see
ZhangLabWiki:Copyrights
for details).
Do not submit copyrighted work without permission!
Cancel
Editing help
(opens in new window)
Navigation menu
Personal tools
Not logged in
Talk
Contributions
Create account
Log in
Namespaces
Page
Discussion
English
Views
Read
Edit
View history
More
Search
Navigation
Main Page
Current events
Recent changes
Random page
Investigators
Matt Cai
Song Chen
Eric Chu
Dinh Diep
Elizabeth Duong
Shicheng Guo
Alan Fung
Daniel Jacobsen
Blue Lake
Huy Lam
Alice Li
Andrew Richards
Brandon Sos
Chris Wei
Yan Wu
Kun Zhang
Tools
What links here
Related changes
Special pages
Page information