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