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pGP-AAV-syn-jGCaMP7f-WPRE in Mouse (AAV9)

Addgene #104488-AAV9

Data Submitted By
Kyle Lillis
Submission Date
July 12, 2019
Publication Date
July 12, 2019 (modified May 16, 2023)
Abstract
Not provided
This green, calcium sensitive fluorescent protein was imaged using blue ~470nm excitation light and a GFP emission filter. A 10Hz, 4min movie was acquired, during which there were multiple seizures. Since GCaMP7f is very dim at rest, a standard-deviation projection of the entire movie was computed to highlight active cells. This is the image uploaded.
Caption: This green, calcium sensitive fluorescent protein was imaged using blue ~470nm excitation light and a GFP emission filter. A 10Hz, 4min movie was acquired, during which there were multiple seizures. Since GCaMP7f is very dim at rest, a standard-deviation projection of the entire movie was computed to highlight active cells. This is the image uploaded.

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

Vector Used
pGP-AAV-syn-jGCaMP7f-WPRE
Addgene #104488-AAV9

Virus & Injection

Virus

Virus Name
pGP-AAV-syn-jGCaMP7f-WPRE
Serotype
AAV9
Source
Addgene
Catalog Number
104488-AAV9
Promoter
Synapsin
Cargo Type
Biosensor
Injection Titer / Dose
2.50E+10 GC/ml
Injection Volume
Not provided
Injection Rate
Not provided
Injector Material
Not provided
Injection Site / Route
N/A
Additional Details
Slices were incubated with 5μl of AAV in 2mL GBSS + glucose, bubbled with 95/5 for 1hr after slicing and before placing on membrane. This produces widespread expression in the slice; Hippocampal neurons
View in Addgene Catalog

Other Details

Species
Mouse
Strain or Cell Line
C57/BL6J
Source
Jackson Labs
Stock Number
JAX: 000664
Age at Injection
P7 (DIV0)
Time After Injection
1 week
Detection Method
Direct fluorescence
Assay & Results
Not provided
Functional Testing
Organotypic slices spontaneously develop seizures in culture. This calcium-sensitive fluorescent protein vector reliably reported seizure activity in hippocampal neurons as expected
Other Information
No toxicity was observed
Tips & Comments
AAV9 works well in our hands for injections. When slices are incubated in the virus, as in this experiment, capsid seems to be less important