For mice with CAV-Cre injected to the BLA or mPFC and AAV-DIO-hM3D-mCherry into the vMT, we compared the number of mCherry+ cells in the vMT. For c-Fos screens, we compared the number of c-Fos+ cells in the vMT and Xi in control mice and mice that experienced visual threats. After we immunostained the brains with identical protocols, the brains were imaged on an automated slide scanner (Zeiss). A brain map was overlain on the digital image to identify the appropriate regions using landmark structures, white matter tracts, ventricles and optic tract as reference guides. An experimenter blind to condition analysed the images using the automated ImageJ cell counter.
Automated slide scanner
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2 protocols using automated slide scanner
1
Quantifying Cell Activation in Mouse Brain
2
Quantifying vLGN Projections and Labeling
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For mice with injections to the vLGN, we quantified the relative
density of labeled axons in the Re versus Xi portions of the vMT. For mice
with CTβ injections to the mPFC or BLA, we quantified the number of
retrogradely labeled cells within the Re and Xi. For Gad2-IRES-Cre mice with
injections of AAV-DIO-hM3D-mCherry, we quantified the number of labeled
hM3D/mCherry+ cells within the external and internal division of the vLGN.
For mice with injections of AAVretro-EF1a-Nuc-flox(mcherry)-eGFP, we
quantified the number of retrogradely labeled GFP+ and mCherry+ cells within
the vLGN. The brains were imaged on an automated slide scanner (Zeiss). A
brain map was overlain on the digital image to identify the appropriate
regions using landmark structures, ventricles and optic tract as reference
guides. An experimenter blind to condition analyzed the images in ImageJ.
For axon density, the mean fluorescence intensity was measured in the Re and
the Xi portion of the vMT. For cell counts, the number of labeled cells were
analyzing using the automated ImageJ cell counter.
density of labeled axons in the Re versus Xi portions of the vMT. For mice
with CTβ injections to the mPFC or BLA, we quantified the number of
retrogradely labeled cells within the Re and Xi. For Gad2-IRES-Cre mice with
injections of AAV-DIO-hM3D-mCherry, we quantified the number of labeled
hM3D/mCherry+ cells within the external and internal division of the vLGN.
For mice with injections of AAVretro-EF1a-Nuc-flox(mcherry)-eGFP, we
quantified the number of retrogradely labeled GFP+ and mCherry+ cells within
the vLGN. The brains were imaged on an automated slide scanner (Zeiss). A
brain map was overlain on the digital image to identify the appropriate
regions using landmark structures, ventricles and optic tract as reference
guides. An experimenter blind to condition analyzed the images in ImageJ.
For axon density, the mean fluorescence intensity was measured in the Re and
the Xi portion of the vMT. For cell counts, the number of labeled cells were
analyzing using the automated ImageJ cell counter.
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