(2022). eLife, 11, . doi: 10.7554/elife.76254. Article
To directly compare mGreenLantern to mScarlet, adult mice received lumbar (L1-L2) injection of mixed AAV2-retro-H2B-mGreenLantern and -mScarlet, followed 2 weeks later by brain clearing, 3D imaging, and nuclei detection using Imaris software. Quantification of labeled objects revealed that H2B-mGreenLantern significantly increased detection of neurons in cortex, dorsal pons, and reticular formation (p<0.01, two-way ANOVA with post-hoc Sidak’s) (Figure 1—figure supplement 2L–O). Moreover, signal was readily detectable without the need for antibody amplification, thus avoiding lengthy incubation periods. Combined, these data establish an initial categorization and quantification of supraspinal brain regions in 3D space, reveal H2B-mGL to be the most sensitive of the FPs tested, and create consistent experimental parameters for the detection of supraspinal neurons.