AAV Injection into Mouse Visual Cortex

Constructs used to produce AAV included pGP-AAV-SYN1-red GECI-WPRE and the cre-recombinase-activated construct pGP-AAV-FLEX-SYN1-red GECI-WPRE. Virus (titer: ~0.7-3x10¹³ genomes/ml, R-CaMP2 promoter was CaMKII) was slowly injected (25–30 nl over 5 min) through a thinned skull into the primary visual cortex (1–2 injection sites at variable depths; 2.7 mm lateral and 0.2 mm anterior to lambda suture; 250 μm deep for L2/3 imaging, 250 μm and 450 μm for L4 imaging, 500 μm for L5 imaging, and 650 μm and 900 μm for L6 imaging).

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Dana H., Mohar B., Sun Y., Narayan S., Gordus A., Hasseman J.P., Tsegaye G., Holt G.T., Hu A., Walpita D., Patel R., Macklin J.J., Bargmann C.I., Ahrens M.B., Schreiter E.R., Jayaraman V., Looger L.L., Svoboda K, & Kim D.S. (2016). Sensitive red protein calcium indicators for imaging neural activity. eLife, 5, e12727.

Publication 2016

Camkii Cre recombinase Genomes Injection sites Primary visual cortex Skull Suture Syn1 Virus

Corresponding Organization :

Other organizations : Howard Hughes Medical Institute, Janelia Research Campus, Weizmann Institute of Science, Rockefeller University

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Variable analysis

independent variables

Virus injection site (primary visual cortex)
Virus injection depth (250 μm, 450 μm, 500 μm, 650 μm, 900 μm)
Virus injection volume (25–30 nl over 5 min)

dependent variables

Virus titer (~0.7-3x10^13 genomes/ml)
Promoter (CaMKII)

control variables

Injection method (slow injection)
Injection site location (2.7 mm lateral and 0.2 mm anterior to lambda suture)

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