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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: Tracking distinct microglia subpopulations with photoconvertible Dendra2 in vivo

Fig. 4

Photoconversion is stable for several days and can differentiate subpopulations of microglia in healthy and degenerating retina. A In healthy, dark reared Arr1−/− Cx3cr1+/D2 mice, 24 h after a large portion of retina was photoconverted in vivo (150 µW, 780 × 780 µm, 6 J/cm2 radiant exposure), 16% of flow cytometry-isolated cells were rD2 positive. B, C Similar numbers were found in degenerating Arr1−/− Cx3cr1+/D2 retina after 24 h B and 48 h C of light exposure, demonstrating stability of the photoconverted microglia population over multiple days in degenerating retina. D In degenerating retinas that were not subjected to photoconversion, only 1–2% of cells were rD2 positive, which is similar to unconverted controls in healthy retina. Error bars are SE, n = 2 retinas, 1 male/1 female for each experimental condition. EG In Arr1−/− Cx3cr1+/D2 (KO) and Arr1+/+ Cx3cr1+/D2 (WT control) mice, resident microglia in the imaging area were photoconverted (6 J/cm2) prior to light exposure. Comparison of pre-degenerating E and degenerating F retina in the same mouse revealed that after 24 h of light exposure, bright green cells that had not been photoconverted were visible within and near retinal vessels only in degenerating retina (F, white arrows in zoomed images) and were clearly distinguishable from resident microglia that appeared yellow/orange and contained gD2 and rD2. Similar bright green, single labeled cells were not observed in Arr1 wildtypes G. Images E, F from same female mouse, G from a male mouse; panels to the right of F are digitally magnified images from the dashed box area in the full imaging field to the left; white lines denote vasculature in zoomed images. Widefield scale 300 µm, zoomed scale 100 µm

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