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UND 363 Oil Red O
| Question | Answer |
|---|---|
| Purpose for Oil Red O stain | demo neutral lipids in FS tissue, eval normal and abnormal fat tissue, fat tumor cells from other tumor cells |
| Principle for Oil Red O stain | staining with oil soluble dyes (ones that are better soluble with lipoids) Vs. usual hydroalcoholic dyes |
| general principles for Oil Red O stain | lipids are soluble in standard histo processes (therefore FS or fixed must be used), small droplets of fat are less likely to be dissolved in propylene glycol than alcohol. |
| what can Oil Red O be dissolved in | can be dissolved in propylene glycol rather than isopropanol |
| what typ of staining method is oil red | physical |
| what are the 4 things the dye use must be | more soluble in tissue lipid than in the solvent it is dissolved in, not be water solube, strongly colored, act with tissue only by solution |
| what is the difference in isopropanol and propylene glycol in relation to lipid | isoprop removes a minimal amount of lipid while prop.glycol will not extract any |
| fix for Oil Red O stain | 10% NBF or calcium formol (bouins ok as well) |
| what fixative should not be used | NO ALCOHOLIC FIXES |
| tech for Oil Red O stain | FS at 10 microns (dehydrating and clearing agents dissolve fat) |
| QC for Oil Red O stain | no control needed, most tissue contains fat |
| what is the first part of the procedure and some special requirements | cut FS, fix in 40% formaldehyde for 1 min. |
| reagents for (in order) | 40% formaldehyde fix FS first... then, oil red o - physical fat stain, harris hematox - nuclei stain, ammonia h20- blue sections, aqueous mount media (must seal edges with nail polish) |
| results for Oil Red O stain | fat - red, nuclei - black |
| what are lipids removed by and therefore should be avoided | fix containing alcohol or organic solvents |
| what are the preferred sections for this stain | free floating sections stain better but are difficult to get with cryostat |
| what can be done to improve microtomy of FS sections | formalin fix tissue can be infiltrated with 30% sucrose before freezing |
| what mount media must be used and what should one be careful of if glycerin jelly is used for mount | aqueous mout media must be used (synthetics will dissolve fat), if glycerin jelly do not overheat or it will melt fat and displace it. |
| what is a concern with coverslipping | fat is mobile, threfore don't put any pressure on the cover glass or the fat will be displaced |
| if oil red o is dissolved in propylene glycol what temp should be used | 60○C (isopropanol is more likely to dissolve small fat droplets) |
Created by:
mustangvxd
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