title: FluoroMyelin™ Red is a bright, photostable and non-toxic fluorescent stain for live imaging of myelin.
abstract: FluoroMyelin™ Red is a commercially available water-soluble fluorescent dye that has selectivity for myelin. This dye is marketed for the visualization of myelin in brain cryosections, though it is also used widely to stain myelin in chemically fixed tissue. Here we have investigated the suitability of FluoroMyelin™ Red as a vital stain for live imaging of myelin in myelinating co-cultures of Schwann cells and dorsal root ganglion neurons. We show that addition of FluoroMyelin™ Red to the culture medium results in selective staining of myelin sheaths, with an optimal staining time of 2h, and has no apparent adverse effect on the neurons, their axons, or the myelinating cells at the light microscopic level. The fluorescence is bright and photostable, permitting long-term time-lapse imaging. After rinsing the cultures with medium lacking FluoroMyelin™ Red, the dye diffuses out of the myelin with a half life of about 130 min resulting in negligible fluorescence remaining after 18-24h. In addition, the large Stokes shift exhibited by FluoroMyelin™ Red makes it possible to readily distinguish it from popular and widely used green and red fluorescent probes such as GFP and mCherry. Thus FluoroMyelin™ Red is a useful reagent for live fluorescence imaging studies on myelinated axons.
authors: Department of Neuroscience, The Ohio State University, Columbus, OH 43210, United States.
authors: Paula C
authors: PC
authors: Monsma
authors: Anthony
authors: A
authors: Brown
mesh_terms: D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D004396:Coloring Agents / Q000378:metabolism; D004622:Embryo, Mammalian; D005260:Female; D005456:Fluorescent Dyes / Q000378:metabolism*; D005470:Fluorometry; D005727:Ganglia, Spinal / Q000166:cytology; D049452:Green Fluorescent Proteins / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D018993:Myelin P0 Protein / Q000378:metabolism; D009186:Myelin Sheath / Q000378:metabolism*; D009474:Neurons / Q000502:physiology; D011247:Pregnancy; D051381:Rats; D012583:Schwann Cells / Q000378:metabolism*; D013997:Time Factors; D014162:Transfection; D000097573:Red Fluorescent Protein
chemical_list: D004396:Coloring Agents; D005456:Fluorescent Dyes; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D018993:Myelin P0 Protein; D000097573:Red Fluorescent Protein; C485184:enhanced green fluorescent protein