Retinal Cell Identification Protocol

After recording, retinas were fixed (4% paraformaldehyde, 30 min, 20°C) and counterstained with one or more antibodies: (1) rabbit anti-cocaine and amphetamine-regulated transcript (CART; 1:1000; H00362, Phoenix Pharmaceuticals), a specific marker for most ON-OFF DS cells; Kay et al., 2011 (link)); (2) guinea pig anti-RNA-binding protein with multiple splicing (RBPMS; 1:1000; 1832-RBPMS, PhosphoSolutions), a pan-ganglion-cell marker; Rodriguez et al., 2014 (link)); (3) goat anti-ChAT (1:100, anti-ChAT; AB144P, Millipore); (4) rat anti-mCherry (1:1000, EST202, Kerafast); or (5) rabbit anti-melanopsin (1:1000, Advanced Targeting Systems).

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Sabbah S., Berg D., Papendorp C., Briggman K.L, & Berson D.M. (2017). A Cre Mouse Line for Probing Irradiance- and Direction-Encoding Retinal Networks. eNeuro, 4(2), ENEURO.0065-17.2017.

Publication 2017

H00362 1832-RBPMS AB144P

Amphetamine Antibodies Cart Cell Cocaine Ganglion Goat Guinea pig Melanopsin Paraformaldehyde Pharmaceuticals Rabbit Rbpms Retinas

Corresponding Organization :

Other organizations : John Brown University, National Institutes of Health

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Variable analysis

independent variables

Antibody type (rabbit anti-CART, guinea pig anti-RBPMS, goat anti-ChAT, rat anti-mCherry, rabbit anti-melanopsin)

dependent variables

Presence and distribution of specific cell types (ON-OFF DS cells, ganglion cells, cholinergic cells, mCherry-expressing cells, melanopsin-expressing cells)

control variables

Fixation method (4% paraformaldehyde, 30 min, 20°C)
Temperature (20°C)

controls

Positive control: None explicitly mentioned
Negative control: None explicitly mentioned

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