AAV pCAG-FLEX-tdTomato-WPRE in Mouse (AAV9)
Addgene #51503-AAV9

This image was captured using a custom inverted single-photon widefield fluorescence microscope (shown is the red channel only). An Olympus 20X 0.95 NA objective was used to acquire a 5x5 grid of z-stacks. The image shown is a stitched maximum z-projection spanning essentially the entire volume of the slice.

Caption: This image was captured using a custom inverted single-photon widefield fluorescence microscope (shown is the red channel only). An Olympus 20X 0.95 NA objective was used to acquire a 5x5 grid of z-stacks. The image shown is a stitched maximum z-projection spanning essentially the entire volume of the slice.

Viruses & Injection

Primary Virus
Promoter
CAG
Injected Titer
1.00E+13 GC/ml
Injected Volume
5 μl
Injection Rate
Unknown, manual injection
Injection Site
Cerebrospinal fluid (CSF): Intracerebroventricular (ICV)
View in Addgene Catalog
Secondary Virus
pGP-AAV-syn-jGCaMP7f-WPRE (AAV9) was co-injected but is not shown in the image. (AAV9)
Promoter
Syn
Injected Titer
1.00E+13 GC/ml
Injected Volume
5 uL
Injection Rate
Unknown (manual)
Injection Site
Cerebrospinal fluid (CSF): Intracerebroventricular (ICV)
View in Addgene Catalog

Subject

Species / Strain
Mouse Dlx5/6-cre mice from Jackson Labs. Strain 008199
Stock Number
8199
Age at Injection
P1

Data Collection

Time After Injection
1 week
Detection Method
Direct fluorescence
Expression Site
Imaging was performed on hippocampal organotypic slice cultures. Interneurons expressed the construct
Genetic Tool Outcome
The tool was intended to generate red fluorescence in all dlx-cre-expressing interneurons. The pattern of expression matched that expected and was very bright
Tips & Comments
ICV injections at P0-P1 of AAV9 works very well at producing widespread expression throughout the brain, particularly in the hippocampi. Vectors with the CAG promoter make for very bright expression - I would probably use a smaller volume of AAV in the future, for the sake of minimizing the impact of injection