Daniel:Notebook/RNAFACS/2015-10-23: Difference between revisions

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=RNA Scope Trial 5=
=RNA Scope Trial Run 5: Pretreat II (Started [[Daniel:Notebook/RNAFACS/2015-10-22|10/22/2015]])=


[[Daniel:Notebook/RNAFACS|Back to Calendar]]
[[Daniel:Notebook/RNAFACS|Back to Calendar]]
==Microscopy Results==
Here are the microscopy results from the trial today:
<gallery heights=300px widths=300px perrow=3 caption="Taken with 63X oil on confocal; max z-plane; Red=Draq5 (C2), Green=FEZF2 (C3); Yellow=RBFOX3/NeuN (C1)">
File:20151023_NeuN_FEZF2.lif_Field1_Pack_ch00.jpg|Field 1; line average not taken
File:20151023_NeuN_FEZF2.lif_Field2_LineAverage_ch00.jpg|Field 2; Line Average=4
File:20151023_NeuN_FEZF2.lif_Field3_ch00.jpg|Field3; Line Average=4
</gallery>
For future use: using line average (when signal is strong) can reduce the amount of background noise seen.  The field 1 image has the most background and the highest threshold (~50 for the Green/Yellow), whereas field 2 and field 3 only had around 20 as the intensity threshold (same gain, 750, for all fields/channels).
This is a good result.  The field 1 and field 2 images show some nuclei with both FEZF2 and NeuN signal, whereas field 3 only has NeuN.  We expect to see a large number of nuclei with signal in NeuN and fewer with FEZF2.
[[Category:RNAScope]] [[Category:Microscopy]] [[Category:RNAFACS]] [[Category:PretreatII]]

Latest revision as of 18:37, 27 October 2015

RNA Scope Trial Run 5: Pretreat II (Started 10/22/2015)[edit]

Back to Calendar

Microscopy Results[edit]

Here are the microscopy results from the trial today:

For future use: using line average (when signal is strong) can reduce the amount of background noise seen. The field 1 image has the most background and the highest threshold (~50 for the Green/Yellow), whereas field 2 and field 3 only had around 20 as the intensity threshold (same gain, 750, for all fields/channels).

This is a good result. The field 1 and field 2 images show some nuclei with both FEZF2 and NeuN signal, whereas field 3 only has NeuN. We expect to see a large number of nuclei with signal in NeuN and fewer with FEZF2.