pZac2.1 gfaABC1D-lck-GCaMP6f in Mouse (AAV5)
Addgene #52924-AAV5

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

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

Viruses & Injection

Primary Virus
Promoter
GFAP
Injected Titer
1.20E+13 GC/ml
Injected Volume
300 nl
Injection Rate
50 nl/min
Injection Site
Brain parenchyma
Brain Region
Forebrain: Cerebral cortex (neocortex)
Injection Site Comments
Somatosensory cortex
View in Addgene Catalog
Secondary Virus
AAV9-hSyn-RCaMP1.07
Promoter
Syn
Injected Titer
2.40E+12 GC/ml
Injected Volume
300 nl
Injection Rate
50 nl/min
Injection Site
Brain parenchyma
Brain Region
Forebrain: Cerebral cortex (neocortex)
Injection Site Comments
Somatosensory cortex

Subject

Species / Strain
Mouse C57/BL6J
Age at Injection
8-12 weeks

Data Collection

Time After Injection
3 weeks
Detection Method
Immunohistochemistry, Direct fluorescence
Expression Site
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
Genetic Tool Outcome
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