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. |
Revision as of 16:42, 23 October 2015
RNA Scope Trial Run 5: Pretreat II (Started 10/22/2015)
Microscopy Results
Here are the microscopy results from the trial today:
- 20151023 NeuN FEZF2.lif Field1 Pack ch00.jpg
Field 1; line average not taken
- 20151023 NeuN FEZF2.lif Field2 LineAverage ch00.jpg
Field 2; Line Average=4
- 20151023 NeuN FEZF2.lif Field3 ch00.jpg
Field3; Line Average=4
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.