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=Overload FISH ([[Daniel:Notebook/HiResChrPaint/2014-3-5|Started 3/5/2014]])= [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] ==Report== '''Sample Matrix''' {| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext> |- style="font-size:12pt;font-weight:bold" align="center" | width="65" height="30" | Sample | width="65" | Sample A | width="65" | uL sample A (pmol) | width="65" | Sample B | width="65" | uL sample B (pmol) | width="65" | Sample B | width="65" | uL sample B (pmol) |- style="font-size:12pt" align="center" |style="font-weight:bold" height="30" | V6S3 +V6S2 (A) | V6S3 | 40 (48) | V6S2 | 20 (98) | Unlabeled Cot1 | 2 (2 ug) |- style="font-size:12pt" align="center" |style="font-weight:bold" height="30" | V6S3+Cot1 (B) | V6S3 | 40 (48) | Cot1 | 20 (10) |style="text-decoration:underline;color:#0000FF" align="center" | |style="text-decoration:none" align="center" | |- style="font-size:12pt" align="center" |style="font-weight:bold" height="45" | BSPP Negative Control (D) | BSPP1B | 5 (10) | Unlabeled Cot1 | 2 (2 ug) | align="center" | | align="center" | |} ===Sample C (BSPP1)=== 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. {| class="wikitable" |- |[[Image:20140310_BSPP1_ManyNucleii1.jpg|1000px]]|[[Image:20140310_BSPP1_ManyNucleii2.jpg|1000px]]| |- |Nucleii from BSPP1 sample, 63x objective, 1x zoom. Draq5 in blue, Alexa 488 in green|| |} ===Sample B (V6S3-546 and Cot1-488)=== 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. {| class="wikitable" |- |[[Image:20140310_V6S3Cot1_TwoNucleii.jpg|1000px]]|[[Image:20140310_V6S3Cot1_ThreeNucleii.jpg|1000px]]| |- |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)=== 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. {| class="wikitable" |- |[[Image:20140310_V6S2V6S3_ManyNucleii1.jpg|1000px]]|[[Image:20140310_V6S2V6S3_ManyNucleii2.jpg|1000px]]| |- |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... {| class="wikitable" |- |[[Image:20140310_V6S2V6S3_SingleNucleus.jpg|1000px]]| |- |Nucleii from V6S2-V6S3 sample, 63x objective, 1x zoom. Draq5 in blue, Alexa 488 in green, Alexa 546 in red|| |} [[Category:HRCP]] [[Category:20140305]] [[Category:Microscopy]]
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