Daniel:Notebook/HiResChrPaint/2014-3-13

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Overload FISH (Started 3/5/2014)[edit]

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Report[edit]

Sample Matrix

Sample Sample A uL sample A (pmol) Sample B uL sample B (pmol) Sample B uL sample B (pmol)
V6S3 +V6S2 (A) V6S3 40 (48) V6S2 20 (98) Unlabeled Cot1 2 (2 ug)
V6S3+Cot1 (B) V6S3 40 (48) Cot1 20 (10)    
BSPP Negative Control (D) BSPP1B 5 (10) Unlabeled Cot1 2 (2 ug)    


Sample C (BSPP1)[edit]

This sample is just BSPP1 labeled with A-488. As we expect, there is not a lot of nuclear signal, and it is in no particular patterning.

File:20140310 BSPP1 ManyNucleii1.jpg|File:20140310 BSPP1 ManyNucleii2.jpg|
Nucleii from BSPP1 sample, 63x objective, 1x zoom. Draq5 in blue, Alexa 488 in green

Sample B (V6S3-546 and Cot1-488)[edit]

I think in general this sample was worse than the positive controls I have done previously, but still has clear signal from the labeled cot1 DNA and poor signal from the paint probes.

File:20140310 V6S3Cot1 TwoNucleii.jpg|File:20140310 V6S3Cot1 ThreeNucleii.jpg|
Nucleii from V6S3-Cot1 sample, 63x objective, 1x zoom. Draq5 in blue, Alexa 488 in green, Alexa 546 in red

Sample A (V6S2-488 + V6S3-546)[edit]

The sample images show a lot of signal within the nucleus (and not too much without), but I'd like to do a little post-processing on them to see if more distinct regions can be parsed out.

File:20140310 V6S2V6S3 ManyNucleii1.jpg|File:20140310 V6S2V6S3 ManyNucleii2.jpg|
Nucleii from V6S2-V6S3 sample, 63x objective, 1x zoom. Draq5 in blue, Alexa 488 in green, Alexa 546 in red

And in case the zoom needs to be higher...


File:20140310 V6S2V6S3 SingleNucleus.jpg|
Nucleii from V6S2-V6S3 sample, 63x objective, 1x zoom. Draq5 in blue, Alexa 488 in green, Alexa 546 in red