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O0625

Sigma-Aldrich

Oil Red O

Dye content, ≥75%, certified by the Biological Stain Commission, powder

Synonym(s):

1-([4-(Xylylazo)xylyl]azo)-2-naphthol, 1-[2,5-Dimethyl-4-(2,5-dimethylphenylazo)phenylazo]-2-naphthol, Solvent Red 27

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€47.50
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€113.00

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25 G
€47.50
100 G
€113.00

About This Item

Empirical Formula (Hill Notation):
C26H24N4O
CAS Number:
1320-06-5
Molecular Weight:
408.49
Colour Index Number:
26125
EC Number:
215-295-3
MDL number:
MFCD00003898
UNSPSC Code:
12171500
PubChem Substance ID:
24897931
NACRES:
NA.47

€47.50

Please contact Customer Service for Availability
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Product Name

Oil Red O, certified by the Biological Stain Commission

grade

certified by the Biological Stain Commission

Quality Level

200

form

powder

composition

Dye content, ≥75%

technique(s)

microbe id | staining: suitable

color

dark red to brown

mp

120 °C (dec.) (lit.)

solubility

chloroform/ethanol (1:1): 1 mg/mL

application(s)

diagnostic assay manufacturing
hematology
histology

storage temp.

room temp

SMILES string

Cc1ccc(C)c(c1)\N=N\c2cc(C)c(cc2C)\N=N/c3c(O)ccc4ccccc34

InChI

1S/C26H24N4O/c1-16-9-10-17(2)22(13-16)27-28-23-14-19(4)24(15-18(23)3)29-30-26-21-8-6-5-7-20(21)11-12-25(26)31/h5-15,31H,1-4H3

InChI key

NPGIHFRTRXVWOY-UHFFFAOYSA-N

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General description

Oil Red O is a lysochrome.[1] It is used to stain and visualise general localization of fats.[2]
Oil Red O stains lipid materials and the lipids take red-orange color. It also marks vacuoles, particularly observed in ALL-L3 (acute lymphoblastic leukemia) or Burkitt′s lymphoma/leukemia. Oil Red O staining is done on fresh samples, since alcohol fixation removes lipid.[3][4]

Application

Oil Red O has been used for the staining of lipid droplets and lipid-containing vacuoles in cells.[5][6]
Oil Red O has been used:
  • for staining liver sections in histological analysis[7][8][9]
  • for the quantification of lipids in cells[10][11][12]
  • to stain aorta to examine atherosclerotic lesions[13]

Suitability

Certified for use as a fat stain in Churukian′s modification of the method of Lillie and Asburn.

Storage Class Code

11 - Combustible Solids

WGK

WGK 3

Personal Protective Equipment

dust mask type N95 (US), Eyeshields, Gloves

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Certificates of Analysis (COA)

Lot/Batch Number
0000352489
SHBR5465
SHBR9443
SHBR3449
0000352489

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PARP1-mediated PPARalpha poly (ADP-ribosyl) ation suppresses fatty acid oxidation in non-alcoholic fatty liver disease
Huang K, et al.
Journal of Hepatology, 66(5), 962-977 (2017)
Natural dyes versus lysochrome dyes in cheiloscopy: A comparative evaluation
Singh NN, et al.
Journal of forensic dental sciences, 2(1), 11-11 (2010)
DJ-1 Deficiency Protects Hepatic Steatosis by Enhancing Fatty Acid Oxidation in Mice
Xu M, et al.
International Journal of Biological Sciences, 14(13), 1892-1892 (2018)
Adult bone marrow is a rich source of human mesenchymal 'stem' cells but umbilical cord and mobilized adult blood are not.
Wexler SA, et. al.
British Journal of Haematology, 121, 368-368 (2003)
Clinical Laboratory Medicine (2002)
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Questions

1–10 of 11 Questions  

  1. Can this Oil Red O be utilized for Oval Fat Body identification in urinalysis and if so how would you prepare the solution for this usage.

    1 answer

    1. No specific Oil Red O staining procedures are available and validated for staining urine sediment. Below are instructions for preparing the stock and working solutions for use in staining tissue sections. Preparing the stock and working solutions should be the same as if staining tissue sections. It may be necessary to dry the urine sediment onto the slide before use. If the urine sediment has not dried, it is possible the lipid might get washed off the slide during the staining procedure. Do not attempt to fix the urine sediment to the slide using alcohol. If fixation is required, use 10% Neutral buffered formalin as it is considered an aqueous fixative.

      Below are instructions for staining tissue sections. If the urine sediment is attached to the glass slide, the same procedure should be suitable for urine sediment.

      Stock Solution.
      Prepare a saturated (approximately 1%) stock solution of oil red O in 99% isopropanol. Mix well to dissolve.

      Working Solution
      Mix 6 parts of stock solution with 4 parts of deionized water. Let stand for 10 minutes and filter. Working solution stable for 1 - 2 hours.

      Technique
      1. If the urine sediment has been fixed to the slides with 10% Neutral Buffered Formalin, gently rinse sections in deionized water.
      2. Stain in working oil red O solution for 6 - 15 minutes.
      3. Clear background if necessary using 60% isopropanol.
      4. Wash in deionized water.
      6. Coverslip using an aqueous mounting medium or simply use water for the coverslipping.
      Results
      Lipids - orange to red

      It is also possible to view lipids in urine sediment by using polarized microscopy. Another option might be to use a method for fecal fats since the urine sediment would not need to be attached to the glass slide prior to staining.

      Helpful?

  2. If the stock is prepared with chloroform/ethanol (1:1) , what should be used for dilution to working concentration, and what is the staining protocol for staining adipocytes in cell culture?

    1 answer

    1. The most common solvent to dissolve Oil Red O are 99% isopropyl alcohol or propylene glycol. There are no recommendations dissolving the product in a mixture of chloroform/ethanol. When the product is sent to the Biological Stain Commission for certification, the BSC tests staining performance using the Lillie and Ashburn (1943) method as modified by Churukian. The solvent used by the BSC is 99% isopropanol. The BSC prepares a 1% saturated solution in 99% isopropanol. Other sources recommend using a dye content of 0.3% dye content for preparing the stock solution. Working solutions are typically prepared by diluting 6 parts of the Oil Red O stock solution with 4 parts 1% dextrin solution. Most standard protocols use only water. The Churukian modification calls for diluting the stock Oil Red O solution in 1% dextrin. Stock solutions diluted in dextrin are stated to be stable for several weeks. After the working solution is prepared, let stand 10 minutes and filter. The working solution diluted in water are typically stable for only 1-2 hours.

      Cultured adipocytes in cell culture are stained in the same fashion as frozen sections attached to glass slides. The tissue/cells should be first fixed in 10% Neutral Buffered formalin. Avoid alcohol or solvent based fixatives as they will extract fat or lipids from the tissue/cells. Tissues/Cells are than stained for 10 minutes or longer and then rinsed in water. The tissue/cells can then be counterstained with Mayers Hematoxylin, rinsed in water (the nuclei turn blue) and again rinsed in water. The tissue/cells can then be coverslipped using an aqueous mounting media - such as product GG1 or Kaisers Glycerol Jelly

      Helpful?

  3. Does Oil Red O (O0625) stain non polar lipids?

    1 answer

    1. There is no information available except that Oil Red O stains fats or lipids.

      Helpful?

  4. How to prepare the solution without dextrin to stain cell culture hepatocytes?

    1 answer

    1. To stain tissue without the use of dextrin, simply prepare a saturated solution of Oil Red O at 0.5% in isopropanol or ethanol. Immediately before staining, dilute 4 parts of the Oil Red O stock solution with 6 parts water. Let stand for 10 minutes. Filter before use. Working solution without dextrin should be stable 1-2 hours.

      Helpful?

  5. How is the powder form mixed? Which solutions do I need in order to stain my slides. I thought I was ordering the liquid, but ordered the powder by mistake.

    1 answer

    1. There are several methods to use Oil Red O (O0625) solution for lipid detection in frozen sections. We recommend the following one:
      Prepare a saturated (approximately 1%) stock solution of oil red O in 99% isopropanol.
      Dilute 60 ml of stock solution with 40 ml 1% dextrin. Any type of dextrin is suitable.
      Let stand overnight and then filter. The filtrate, which is the working solution, can be used for several weeks.
      Frozen sections are stained 10 minutes or longer, rinsed in 60% isopropanol and then in water.
      Nuclei are counterstained for 5 minutes in an acid alum hematoxylin of about 0.1% strength (e.g., Mayer's undiluted, Lillie's diluted 1 to 4 in 2% acetic acid, or Ehrlich's diluted 1 to 5 in 2% acetic acid) then placed in 0.3% sodium borate or in tap water, until blue.
      Sections are mounted in a suitable aqueous medium such as glycerin jelly, Apathy's syrup, Zwemer's glychrogel or Kaiser's mounting medium.

      Results: Fat globules orange-red; nuclei blue.
      Working solutions prepared without dextrin typically have shorter stability dating.

      For a proper account of this method, read this paper: Churukian, C 1999 Lillie's oil red O method for neutral lipids. J. Histotechnol. 22:309-311.
      The method of the Biological Stain Commission is listed in: Penny, DP et al, Analysis and testing of biological stains - The Biological Stain Commission Procedures. Biotech. Histochem. 77(5&6), 237-275, (2002).
      The Churukian method is also listed in Bancroft, J and Gamble, M; Theory and Practice of Histological Techniques, Churchill Livingstone, New York 2002 pages 207-208.

      Helpful?

  6. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

      Helpful?

  7. Can Oil Red O, Product O0625, be utilized for staining adipocytes in cell culture?

    1 answer

    1. Yes, although the cultured cells must first be fixed before staining.  Preferred fixatives would include aqueous fixatives such as 10% neutral buffered formalin.  Avoid fixatives such as ethanol or acetone. After fixation and rinsing, the cells may be stained with the basic procedures used for staining tissues attached to glass slides

      Helpful?

  8. Has product O0625, Oil Red O, been use-tested for staining lipoproteins in agarose gels?

    1 answer

    1. Each lot of Product O0625 has been certified for use in histological applications by the Biological Stain Commission. It is not specifically use-tested for any electrophoresis application. We offer a different grade of Oil Red O (Product No. O9755) which has been tested for suitability in electrophoresis; each lot of O9755 has been tested and found suitable for staining lipoproteins that have been separated by cellulose acetate electrophoresis.

      Helpful?

  9. Can the Oil Red O, Product O0625, staining procedure be used with paraffin sections of tissue?

    1 answer

    1. No.  The solvents typically used in paraffin processing will dissolve away most of the fats and lipids.  Frozen sections are the preferred tissue specimen.

      Helpful?

  10. Is there a recommended staining procedure for Product O0625, Oil Red O, for staining fats or lipids in tissue sections.

    1 answer

    1. Yes, there are several.  Working solutions prepared without dextrin typically have shorter stability dating.  The method of the Biological Stain Commission is listed in: Penny, DP et al, Analysis and testing of biological stains - The Biological Stain Commission Procedures. Biotech. Histochem. 77(5&6), 237-275, (2002).  The Churukian method is also listed in Bancroft, J and Gamble, M; Theory and Practice of Histological Techniques, Churchill Livingstone, New York 2002 pages 207-208.

      Helpful?

1–10 of 11 Questions  

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