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UND 363 Luxol/Holmes
UND 363 Luxol blue/Holmes Silver
| Question | Answer |
|---|---|
| purpose | demo Myelin AND nerve fibers |
| holmes principle | alkaline impregnation soln' (boric acid, soduim borate, silver nitrate, pyridine) and a reducing solution (hydroquinone and sodium sulfate) |
| luxol fast blue principle | stains due to lipoproteins. it is an adic-base reaction that forms salt. the base of lipoprotein replaces the base of the dye. |
| holmes procedure | mpregnate with alkaline soln, reducer (hydroquinone/soduim sulfite), tone with gold chloride, oxalic acid (reduces gold chloride and gives purple tinge), soduim sulfate to remove excess silver) |
| luxol fast procedure | luxol fast blue (alcohol/sulfonated copper stain), lithium carbonate (diff'r), 70% alcohol (diff'r), |
| fix | formalin |
| equip | chem cleaned glassware |
| tech | 10-15 |
| qc | cerebral cortex (DO NOT use spinal cord because axons will be in cross section) |
| reagents and procedure step by step | impregnation (silver nitrate/boric acid/sodium borate/pyridine), gold toned, oxalic acid (gold reducer and stain intensifier), sod. thio. (remove excess silver), luxol fast (stain myelin) lithium carb then alcohol (diff'rs) |
| results | myelin sheath - blue to green, axon and nerve - black |
| what does the luxol stain and what does the holmes stain | luxol for myelin, and holmes is silver impregnation for nerves |