Optimized multiplex immunofluorescent protocols for simultaneous in situ identification of α-motoneuron subtypes

Key Points

• This research aims to develop a multiplex immunofluorescent protocol for better identification of α-motoneuron subtypes in the spinal cord.
• Novel multiplex protocol designed for labeling four major α-MN types: S, FR, FI, FF.
• Utilized osteopontin, SK2, and SK3 to differentiate α-MN subtypes.
• Applied refined immunofluorescence protocols to mouse lumbar spinal cord.
• SK2 and SK3 successfully label 99% of α-motoneurons.
• The multiplex protocol enables simultaneous labeling of four distinct α-MN types.
• Enhanced specificity in identifying α-MN populations is achieved.