Evaluation of intracellular lipids by standardized staining with a Sudan black B fraction

Subramaniam, Heman N.; Chaubal, K.A.

doi:10.1016/0165-022x(90)90040-j

Cited by 26 publications

(14 citation statements)

References 5 publications

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“…SBB is known to stain all kinds of lipids, including lipofuscin granules. Commercial SBB is mostly composed of two isomeres, SBB-I (20%) and SBB-II (60%), whereas the remaining fraction contains all kinds of impurities (Lansink 1968;Pfüller et al 1977;Subramaniam and Chaubal 1990). SBB has been suggested not to be a true dye (Kiernan 1981) forming a stable chemical bond with lipids but to stain fat because it is more soluble in fat than in the solvent ethanol from which it is to be applied.…”

Section: Blocking Of Autofluorescence With Sudan Black Bmentioning

confidence: 99%

Double Immunolabeling of Neuropeptides in the Human Hypothalamus as Analyzed by Confocal Laser Scanning Fluorescence Microscopy

Romijn

Uum

Breedijk

et al. 1999

J Histochem Cytochem.

132

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SUMMARYThe main goal of this study was to develop a better light microscopic procedure for quantitative study of the cellular co-localization of neuropeptides in adult human brain tissue. To reach this goal, we opted for a method (proved to be optimal on rat brain) in which sections were double immunolabeled with two different fluorophore-conjugated secondary antibodies and analyzed with a confocal laser scanning fluorescence microscope. One of our main problems faced was a strong autofluorescence of the sections, mainly caused by lipofuscin granules normally present in adult human brain tissue, which made any analysis of specific fluorescence impossible. This problem could be solved by staining the sections after immunolabeling with the dye Sudan Black B, which completely blocked this autofluorescence. The complete optimized procedure that we eventually developed can be summarized as follows. After a relatively short fixation time (6-14 days) in 4% freshly depolymerized paraformaldehyde, the resected brain tissue can best be stored in a 30% sucrose solution supplemented with 0.05% NaN 3 at 4C. Stored under these conditions, cryosections from the tissue still reveal good histology and allow successful immunocytochemical staining after a period of 6 months. Double immunolabeling is done by incubating cryo-or paraffin sections in a mixture of two primary antibodies directed against the targeted antigens, followed by incubation with two different fluorophore-conjugated secondary antibodies. Amplification with a biotinylated secondary antibody followed by fluorophore-conjugated streptavidin is possible. Finally, the sections are stained with Sudan Black B, mounted in plain 80% Tris-buffered glycerol, and studied by confocal laser scanning fluorescence microscopy. Sections processed in this way are well suited for qualitative and quantitative analyses of co-localized neuropeptides in human brain tissue.

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Section: Blocking Of Autofluorescence With Sudan Black Bmentioning

confidence: 99%

Double Immunolabeling of Neuropeptides in the Human Hypothalamus as Analyzed by Confocal Laser Scanning Fluorescence Microscopy

Romijn

Uum

Breedijk

et al. 1999

J Histochem Cytochem.

132

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“…Moreover, lipid and calcium moieties of the native proteins were stained by mean of isoelectrophoresis using Sudan Black B and Alizarin Red S respectively as suggested earlier. 17,18 Electrophoretic localization of in-gel enzyme activity:…”

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confidence: 99%

Impact of heavy metals on Oreochromis niloticus fish and using Electrophoresis as Bio-indicator for environmental pollution of Rosetta branch, River Nile, Egypt

Mohamed¹,

Ahmed²,

Fathy³

et al. 2020

ECB

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“…Lipids in liver tissue were detected by SBB staining. [ 15 ] Briefly, frozen sections with 8–15 μm in thickness were washed by distilled water and 70% ethanol. The sections were then immersed into SBB dye solution (saturated SBB in 70% ethanol) for 10–35 min, followed by differentiation in 70% ethanol.…”

Section: Methodsmentioning

confidence: 99%

Amelioration of intestinal barrier dysfunction by berberine in the treatment of nonalcoholic fatty liver disease in rats

Zheng

et al. 2017

Phcog Mag

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Objective:To investigate the effect of berberine (BBR) on intestinal barrier function in nonalcoholic fat liver disease (NAFLD) in rats.Materials and Methods:Rats were divided into three groups: normal diet group (control group [CON group]), high-fat diet feeding group (HFD group), and HFD with BBR group. After 8 weeks of HFD feeding, rats in the BBR group were given BBR intragastrically at a dose of 150 mg/kg daily for 4 weeks. The same volume of normal saline was given to the CON and HFD groups. Liver index was detected, and Sudan black B staining was used to study fatty degeneration, also the expression level of occluding and intestinal flora was analyzed.Results:BBR administration significantly reduced HFD-induced increase in body weight (CON group: 379.83 ± 61.51 g, HFD group: 485.24 ± 50.15 g, and BBR group: 428.60 ± 37.37 g). It obviously alleviated the HFD-induced liver fatty degeneration and histopathological changes of intestinal mucosa according to liver index low-density lipoprotein-cholesterol, high-density lipoprotein-cholesterol, and total cholesterol (P < 0.05). The triglyceride, alanine transaminase, and aspartate aminotransferase levels were greatly elevated after BBR treatment (P < 0.05); while endotoxin, intestinal fatty acid-binding protein, and tumor necrosis factor-α were significantly reduced (P < 0.05). Moreover, we found that BBR could obviously elevate the level of occludin and decrease the level of Faecalibacterium prausnitzii and upregulate the level of bacteroides.Conclusion:BBR provides significant protection in NAFLD through ameliorating intestinal barrier function.SUMMARY Berberine (BBR), an alkaloid that can be isolated from many plants, has been medically used for its wide range of antimicrobial and anti-inflammatory effectsThis is a study of BBR on liver function and intestinal barrier function in nonalcoholic fat liver disease (NAFLD)BBR treatment for NAFLD could significantly restore the liver function and provide significant protection in NAFLD through ameliorating intestinal barrier function. Abbreviations used: BBR: Berberine, NAFLD: Nonalcoholic fat liver disease, ALT: Alanine transaminase, AST: Aspartate aminotransferase, TG: Triglyceride, I-FABP: Intestinal-fatty acid-binding protein, IBD: Inflammatory bowel disease.

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Evaluation of intracellular lipids by standardized staining with a Sudan black B fraction

Cited by 26 publications

References 5 publications

Double Immunolabeling of Neuropeptides in the Human Hypothalamus as Analyzed by Confocal Laser Scanning Fluorescence Microscopy

Double Immunolabeling of Neuropeptides in the Human Hypothalamus as Analyzed by Confocal Laser Scanning Fluorescence Microscopy

Impact of heavy metals on Oreochromis niloticus fish and using Electrophoresis as Bio-indicator for environmental pollution of Rosetta branch, River Nile, Egypt

Amelioration of intestinal barrier dysfunction by berberine in the treatment of nonalcoholic fatty liver disease in rats

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