pGP-AAV-syn-jGCaMP7f-WPRE in Mouse (AAV9)
Addgene #104488-AAV9

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.

Virus & Injection

Primary Virus
Promoter
Syn
Injected Titer
2.50E+10 GC/ml
Injection Site
Incubated organotypic slices in virus at the time of slicing
Injection Site Comments
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
View in Addgene Catalog

Subject

Species / Strain
Mouse C57/BL6J
Stock Number
JAX: 000664
Age at Injection
P7 (DIV0)

Data Collection

Time After Injection
1 week
Detection Method
Direct fluorescence
Expression Site
Hippocampal neurons
Genetic Tool Outcome
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