Oil Red O Stain Kit (Lipid Stain) (ab150678)

Reviews (1)Q&A (3)References (18)

Overview

  • Product name

    Oil Red O Stain Kit (Lipid Stain)
    See all Lipids kits

  • Product overview

    Oil Red O (Lipid Stain) kit is intended for use in the histological visualization of fat cells and neutral fat. This kit may be used ONLY on frozen tissue sections, fresh smears, or touch preps. 

    If you prefer to prepare your own Oil Red O solution, we recommend our solid Oil Red O stain ab146295.


    Oil Red O staining protocol summary:
    - prepare fresh or frozen tissue sections
    - incubate slide in propylene glycol for 2 min
    - incubate slide in oil red o solution for 6 min
    - differentiate section in 85% propylene glycol for 1 min
    - rinse slide twice in water
    - incubate in hematoxylin for 1-2 min
    - rinse slide three times in water
    - coverslip with an aqueous mounting medium


    Other products for staining tissue sections


    Find more kits and reagents in the special stains guide, or products for antigen retrieval, blocking, signal amplification, visualization, counterstaining, and mounting in the IHC kits and reagents guide.

  • Notes

    Oil Red O is a fat-soluble dye that stains neutral triglycerides and lipids. It cannot be used with formaldehyde-fixed paraffin embedded sections as the alcohols used remove most lipids.

    Staining Interpretation

    Fat Cells   Red
    Neutral Fat     Red
    Nuclei Blue

    Control Tissue: Any frozen section containing fat.

Properties

Images

  • ab150678 - Oil Red O Stain Kit (Lipid Stain)
    ab150678 - Oil Red O Stain Kit (Lipid Stain)

    ab150678 Oil Red O Stain Kit (Lipid Stain) staining frozen human human adipose.

  • ab150678 - Oil Red O Stain Kit (Lipid Stain)
    ab150678 - Oil Red O Stain Kit (Lipid Stain)

    ab150678 (Oil Red O stain) staining lipid deposits (red) in frozen normal human skin. The dermis is unstained, whereas the hypodermis is stained, demonstrating dye specificity. Hematoxylin (blue) is used as a counterstain.  

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre.

  • ab150678 - Oil Red O Stain Kit (Lipid Stain)
    ab150678 - Oil Red O Stain Kit (Lipid Stain)

    Oil Red O staining of 3T3-L1 and adipocyte-like cells

    During adipocyte differentiation, cells accumulate lipid. Oil Red O staining kit (ab150678) was used to visualize lipid in 3T3-L1 cells (left) and adipocyte-like cells (right). As expected, lipid accumulates in 3T3-L1 cells after differentiation into adipocyte-like cells.

  • ab150678 - Oil Red O Stain Kit (Lipid Stain)
    ab150678 - Oil Red O Stain Kit (Lipid Stain)

    Staining of Frozen Breast Tissue using ab150678 - Oil Red O Stain Kit.

Protocols

References

This product has been referenced in:

  • Chernyshova MP  et al. Systemic and skin-targeting beneficial effects of lycopene-enriched ice cream: A pilot study. J Dairy Sci 102:14-25 (2019). Read more (PubMed: 30447975) »
  • Kaufman S  et al. Roux-en-Y gastric bypass surgery reprograms enterocyte triglyceride metabolism and postprandial secretion in rats. Mol Metab 23:51-59 (2019). Read more (PubMed: 30905616) »
See all 18 Publications for this product

Publishing research using ab150678? Please let us know so that we can cite the reference in this datasheet.

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Submitted Jun 27 2019

Question

Protocol for staining lipids of cultured cells with Oil Red O

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Answer

This reference contains a protocol for staining cultured cells with Oil Red O.

Hope R. G., McLauchlan J. (2000) J. Gen. Virol. 81:1913–1925.

http://jgv.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-81-8-1913#tab2

The following protocol is adapted from the protocol in the paper and uses the ab150678 reagents. You will need paraformaldehyde (PFA). Do not use methanol or acetone for the fixation as they will solubilize the lipid, and do not use formalin instead of PFA as it contains up to 15% methanol.


Adipocyte Detection


1. Prepare solutions and buffers


2. Wash the cells


Take the cells from the incubator and carefully aspirate the medium. Carefully wash the cells with Dulbecco’s PBS, w/o Ca++/ Mg++ .


Note: Do not disrupt the cell monolayer!


3. Fixation of the cells


Carefully aspirate the PBS and transfer the tissue culture dish to a fume hood. Add enough 4% PFA to cover the cell monolayer. Incubate at room temperature for at least 10-20 min.


4. Prepare Oil Red O staining solution


5. Wash the cells


Carefully aspirate the fixation buffer and wash the cell monolayer with distilled water. Carefully aspirate the water and add enough Propylene glycol (85%) to cover the cell monolayer. Incubate at room temperature for 2 min.


6. Add Oil Red O staining solution


Carefully aspirate propylene glycol and add enough Oil Red O staining solution to cover the cell monolayer. Incubate at room temperature for 5-15 min.


7. Wash the cells


Carefully aspirate the Oil Red O staining solution and add propylene glycol for 5 mins.


Aspirate propylene glycol and wash the cell monolayer several times with distilled water until the water becomes clear. Blot the vessel containing the stained cells up side down on a paper towel to remove as much water as possible.


8. Counterstain the cells


Add enough Hematoxylin solution to cover the cellular monolayer. Incubate at room temperature for 1-2 min.


9. Wash the cells


Aspirate the Hematoxylin solution and wash the cell monolayer several times with distilled water until the water becomes clear. Aspirate the water.


10. Analyze the cells


Intracellular lipid vesicles in mature adipocytes stain bright red, whereas the Hematoxylin stained nuclei appear blue-violet.

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Question


I have a question regarding the Oil Red O (Lipid Stain) (ab150678). In the datasheet you describe only the usage for frozen tissue. Is it also possible to use the kit for cultured adipocytes?

If yes do you also have a protocol for the well plate staining? Does the kit have a CE label?

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Answer



I am sorry to confirm that we do not know if our Oil Red O solution will work for live adipocytes. We do not have a protocol or data for this and most of the protocols I have found require alcoholic solutions (ab150678 does not) and a fixation step (also not included in ab150678) for live cells. Below is a link to publications that may shed some light on the matter, also an example protocol is provided.

http://www.jbc.org/content/278/18/15998.long#ref-11



http://www.boneandcancer.org/MOLab%20protocols%20since%2011-2005/F22-Oil%20Red%20O%20Staining%20of%20fat%20cells.htm



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Question

Oil Red O (Lipid Stain) (ab150678)

I am emailing with regards to a kit I have obtained from Abcam, with the catalogue number ab150678. This is a staining kit for tissue histology, however the protocol provided is quite vague and I cannot find further information online. I would like to know further information to perform this stain, including the questions outlined below:

1) Do the frozen tissues require a fixation method prior to staining - for example, in PFA?

2) How are the solutions applied to the slides - for example, are the slides dipped into a coplin jar of the solutions?

3) How much of each solution is applied per slide, and can the solutions be reused? My confusion arises from that 100 tests are supposed to be possible from the volume of solutions provided, however the volumes appear quite low.

Thank you for looking into my questions. If there is a more detailed protocol for this Oil Red O stain that would also be appreciated.

Thanks,

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Answer

The frozen tissues need to be fixed. Fixing in the PFA is fine. However, when fixing the tissue, avoid using reagents that will dissolve the fats.

We would recommend putting about 200 to 300ul of the solution (depending on the size of the specimen) onto the slides, and not to reuse the solution.

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