Motor neurons were back filled with Alexa 488-dextran conjugate (Invitrogen). The fluorescent dextran was mixed with a small volume of distilled water and the viscous paste was loaded into the tip of a thin insect pin. Larvae grown at different temperatures were anesthetized with 0.1% MS222 and stabbed with the loaded needle through the skin and into the right ventral musculature. Larvae were kept in anesthetic for 15 min followed by washes with 10% MMR where they were allowed to recover for 2 h. Then larvae were anesthetized again and fixed with 2% glutaraldehyde in phosphate buffer for 30 min at room temperature. Fixed samples were dissected to expose the spinal cord by removing the skin and muscle on the right side. Exposed spinal cords were mounted in Vectashield mounting medium H1200 (Vector Labs) and confocally imaged with a 20X objective with a Nikon C2 microscope. Labeled cells were counted from at least 10 larvae per treatment.
Alexa 488 dextran conjugate
Manufactured by Thermo Fisher Scientific
The Alexa Fluor 488-dextran conjugate is a fluorescent labeling reagent. It consists of the Alexa Fluor 488 dye molecule covalently attached to dextran, a polysaccharide. The conjugate can be used to label and track the movement of dextran in various biological applications.
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Lab products found in correlation
2 protocols using alexa 488 dextran conjugate
1
Retrograde Labeling of Motor Neurons
2
Retrograde Labeling of Motor Neurons
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Motor neurons were back filled with Alexa 488-dextran conjugate (Invitrogen). The fluorescent dextran was mixed with a small volume of distilled water and the viscous paste was loaded into the tip of a thin insect pin. Larvae grown at different temperatures were anesthetized with 0.1% MS222 and stabbed with the loaded needle through the skin and into the right ventral musculature. Larvae were kept in anesthetic for 15 min followed by washes with 10% MMR where they were allowed to recover for 2 h. Then larvae were anesthetized again and fixed with 2% glutaraldehyde in phosphate buffer for 30 min at room temperature. Fixed samples were dissected to expose the spinal cord by removing the skin and muscle on the right side. Exposed spinal cords were mounted in Vectashield mounting medium H1200 (Vector Labs) and confocally imaged with a 20X objective with a Nikon C2 microscope. Labeled cells were counted from at least 10 larvae per treatment.
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