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pAAV.Syn.GCaMP6f.WPRE.SV40 in Mouse (AAV9)

Addgene #100837-AAV9

Data Submitted By
Sam Benezra and Randy Bruno
Submission Date
July 09, 2019
Publication Date
July 09, 2019 (modified May 16, 2023)
Abstract
Not Provided
Green channel, Sutter MOM, Zeiss 20x 1.0NA. Image shows a maximum intensity projection across 200um in layer 2/3 neurons. In this animal, layer 4 neurons were also labeled, but they are deeper in the brain and are not shown in this image. This image is a maximum intensity projection from 100 microns down to roughly 300 microns. Layer 4 begins at about 400 microns deep.
Caption: Green channel, Sutter MOM, Zeiss 20x 1.0NA. Image shows a maximum intensity projection across 200um in layer 2/3 neurons. In this animal, layer 4 neurons were also labeled, but they are deeper in the brain and are not shown in this image. This image is a maximum intensity projection from 100 microns down to roughly 300 microns. Layer 4 begins at about 400 microns deep.

Any further use of this image requires permission from the lab who submitted the data.

Vector Used
pAAV.Syn.GCaMP6f.WPRE.SV40
Addgene #100837-AAV9

Virus & Injection

Virus

Virus Name
pAAV.Syn.GCaMP6f.WPRE.SV40
Serotype
AAV9
Source
Addgene
Catalog Number
100837-AAV9
Promoter
Synapsin
Cargo Type
Biosensor
Injection Titer / Dose
1.28E+13 GC/mL
Injection Volume
4 sites, 200 nL per site
Injection Rate
30 nL/min
Injector Material
Not Provided
Injection Site / Route
Cerebral cortex, NeuroNames ID: 39
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
C57/BL6J
Source
Not provided
Age at Injection
12 weeks
Time After Injection
2 weeks
Detection Method
Direct fluorescence, 2-photon microscopy. baseline fluorescence was clearly visible in layer 2/3, as well as spontaneous activity
Assay & Results
Baseline fluorescence was clearly visible in layer 2/3, as well as spontaneous activity. At approximately 6 weeks, some cell nuclei were filled with GCaMP. The animal was then discarded.
Functional Testing
Baseline fluorescence was clearly visible in layer 2/3, as well as spontaneous activity.
Other Information
Expression became visible already at 1 week, and GCaMP is known to be toxic if expression is too high.
Tips & Comments
I would use a lower titer.