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pZac2.1 gfaABC1D-lck-GCaMP6f in Mouse (AAV5)

Addgene #52924-AAV5

Data Submitted By
Jillian Stobart
Submission Date
July 16, 2019
Publication Date
July 16, 2019 (modified May 16, 2023)
Abstract
Not Provided
Caption: In vivo two-photon microscope picture. Green: gfaABC1D-lck-GCaMP6f. Magenta: hSyn-RCaMP1.07. 20x objective, zoom 2. Excitation: 990nm. GaAsP PMTs with 520/70 nm and 607/50 nm (red) band pass filters. See also: Stobart et al. Neuron 2018. DOI: 10.1016/j.neuron.2018.03.050 Figure 1C

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

Vectors Used
pZac2.1 gfaABC1D-lck-GCaMP6f
Addgene #52924-AAV5
AAV9-hSyn-RCaMP1.07

Virus & Injection

Virus 1

Virus Name
pZac2.1 gfaABC1D-lck-GCaMP6f
Serotype
AAV5
Source
Addgene
Catalog Number
52924-AAV5
Promoter
GFAP
Cargo Type
Biosensor
Injection Titer / Dose
1.20E+13 GC/ml
Injection Volume
300 nl
Injection Rate
50 nl/min
Injector Material
Not Provided
Injection Site / Route
Somatosensory cortex, NeuroNames ID: 3241
View in Addgene Catalog

Virus 2

Virus Name
AAV9-hSyn-RCaMP1.07
Serotype
AAV9
Source
Unknown
Catalog Number
N/A
Promoter
Synapsin
Cargo Type
Biosensor
Injection Titer / Dose
2.40E+12 GC/ml
Injection Volume
300 nl
Injection Rate
50 nl/min
Injector Material
Not Provided
Injection Site / Route
Somatosensory cortex, NeuroNames ID: 3241

Other Details

Species
Mouse
Strain or Cell Line
C57/BL6J
Source
Not provided
Age at Injection
8-12 weeks
Time After Injection
3 weeks
Detection Method
Immunohistochemistry, Direct fluorescence
Assay & Results
The gfaABC1D-lck-GCaMP6f (AAV5) virus works really well. It spreads nicely after the injection and shows membrane labelling only in astrocytes. We have never detected expression in other cell types.
Functional Testing
The calcium sensor behaves as expected and nice microdomains are clearly visible.
Other Information
We have observed activated microglia and reactive astrocytes by immunohistochemistry when higher virus titers are injected. However, at this concentration, staining was normal.