Viral injections were performed as previously described (Beier et al., 2016 (link); Ma et al., 2020 (link)). Briefly, VSV and lentivirus stock was diluted to final working concentrations (described in the "Results" section) with DMEM (11995073; Fisher Scientific), with Fast Green dye (BP123-10; Fisher Scientific) as the injection marker. Glass capillaries (TW100F-4; World Precision Instruments) were pulled into injection needles with a pipette puller (P-97; Sutter Instruments), and the tips were trimmed to create a ~10 μm opening. The injection needle was mounted into a microelectrode holder connected to a pneumatic PicoPump (World Precision Instruments). For retina injection, 3 dpf larvae were anesthetized in Tricaine (0.013% w/v, AC118000500; Fisher Scientific) and mounted laterally inside the center chamber of a glass-bottom dish (P50G-1.5-14-F; MatTek) with 1.5% low melting-point agarose (BP1360; Fisher Scientific). 0.5 nl of virus solution was injected inside the temporal retina.
Fast green dye
Manufactured by Thermo Fisher Scientific
Fast green dye is a synthetic dye commonly used in laboratory settings. It is a water-soluble, green-colored dye that is often employed as a staining agent in various biological and chemical applications. The core function of fast green dye is to provide a visual contrast or labeling for specific targets or samples during analysis and experimentation.
Automatically generated - may contain errors
3 protocols using fast green dye
1
Viral Injections in Zebrafish Retina
2
Subretinal Electroporation of Atxn3 Plasmids
3
Subretinal Electroporation of Atxn3 Plasmids
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pEGFP vector or pEGFP-ATXN3 WT was diluted in PBS (6 μg/μl) and mixed with fast green dye (Fisher Scientific, Waltham, MA) (0.1%). Atxn3 KO neonatal mouse pups (P1) were subjected to subretinal electroporation of the plasmid solutions as described previously (López-Begines et al., 2018 (link)). Transient transgenic retinas were collected and processed at postnatal day 25–30 for immunohistochemistry as mentioned above.
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