Hello everyone,
I'm required to do a histology experiment to determine fat accumulation after an HFD and to compare it to a normal diet. I planned to utilize red O oil staining, however, the equipment for this experiment was not available at my university. Besides, I am unable to go to other campuses owing to the lockdown, which seems to be taking an extended period of time. Thus, I wonder if H and E may substitute oil red O stains
H&E staining is not a good substitute for Oil Red O. In an H&E stain adipose tissue shows up as largely empty spaces. Furthermore, any small lipid containing vesicles that might be readily apparent with an Oil Red O stain will not be easily distinguishable using H&E. (See Figure 5 of this paper for exampleArticle Quantification of liver fat in mice: Comparing dual-echo Dix...
)
The greater issue is loss of fatty acids during the tissue processing, you will need to use frozen tissue. Paraffin imbedding would loose most of the fats as you rehydrate the slides, stain and then finally dehydrate the tissue again.
There are ways of getting Haematoxylin (and Fuchsin) to stain via use of Bichromate as a mordant but are a bit more complicated (old references but still good....... https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1748-1716.1911.tb01550.x)
Alternative biochemistry approach: you could try to extract your fatty deposits, homogenise the samples and run on a reverse phase TLC plate. Might be able to see a differentiation between samples (still got the staining issue but may be more practical approach).
Pathologists in their routine practice rarely need special stains to detect fatty tissue and almost always rely on HE. And if you are planning to assess fatty tissue in HE I would strongly recommend to do it in paraffin sections because frozen sections could be misleading.
Dear Khaled Benchoula
The lipid will be dissolved with H&E staining therefore I suggest to use sudan or Oil Red dyes.
Good luck
H & E is not a good alternative for demonstration of fat in tissue, instead frozen tissue section or normal fixed tissue section with using oil red or Sudan black you can get better results.
- Badges
- Science topic
- Similar topics
- Chemistry
- Pharmacology
- Medical Pharmacology
- Clinical Pharmacology