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UND 363 Oil Red O

QuestionAnswer
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