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| Targets |
Leishman's stain does not have a specific pharmacological target as it is a histological stain. Its mechanism of action involves differential staining of cellular components based on their chemical properties. The eosin component is an anionic dye that stains basic components (such as cytoplasmic proteins) pink to red. The polychrome methylene blue is a cationic dye that stains acidic components (such as nucleic acids) blue to purple. The combination of these dyes produces the characteristic staining pattern that allows for the differentiation of various cell types. The stain is used in hematology and cytology for blood smear analysis.
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| ln Vitro |
Advice (This is our suggested protocol, which should be adjusted to suit your particular requirements as it simply offers guidance) [1]. 1. Assemble the stock solutions for Sorensen buffer: Solution A is Na2HPO4·2H2O (9.5 g/L); Solution B is KH2PO4 (9.1 g/L). 2. For two minutes, cover the smear with Leishman stain. 3. Pour twice as much Sorensen's buffer (6.8 pH) onto the slide, then stir slowly for fifteen minutes. 4. After five minutes, rinse and dry the smear under running water.
In vitro, Leishman's stain is used for staining blood smears and bone marrow specimens for microscopic examination. The stain differentiates cellular components, allowing for the identification of leukocytes, malaria parasites, and trypanosomes. In hematology, the stain is used for normal blood examinations and for detecting malaria parasites. The stain's ability to differentiate cellular components makes it useful for identifying cell types and abnormalities. It is used in tropical medicine to diagnose parasitic infections. |
| ln Vivo |
Leishman's stain is not used as a therapeutic agent and has no established in vivo pharmacological activity. It is primarily a histological dye used for staining blood smears and bone marrow specimens in research and diagnostic settings. The stain is used in microscopy for the examination of blood cells and for the detection of malaria parasites. As a research tool, Leishman's stain is not administered to animals for therapeutic purposes but rather used ex vivo for staining and analytical applications.
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| Enzyme Assay |
In vitro staining procedures using Leishman's stain involve preparing a working solution from the stock solution or powder. For blood smear staining, a thin blood smear is prepared on a glass slide and air-dried. The smear is fixed with methanol and then stained with Leishman's stain solution for 1-3 minutes. The stain is diluted with buffer or distilled water and allowed to stand for an additional 2-5 minutes. The slide is then rinsed with water, dried, and examined under a microscope. The stain is soluble in methanol. Staining results can be verified by microscopic examination.
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| Cell Assay |
In vitro cell-based assays using Leishman's stain are typically performed on fixed cells or tissue sections rather than live cells. For blood smear analysis, whole blood is spread on a glass slide, air-dried, and fixed with methanol. The fixed smear is then stained with Leishman's stain for 1-3 minutes, followed by dilution with buffer and further incubation. For bone marrow specimens, similar procedures are followed. Stained cells are examined under a light microscope for morphological analysis. The stain's ability to differentiate cellular components makes it useful for identifying cell types and abnormalities.
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| Animal Protocol |
Leishman's stain is not used for in vivo animal experiments as it is a histological stain. However, blood samples may be collected from animals for hematological analysis using Leishman's stain. Bone marrow aspirates from animals can be stained for cytological examination. The stain is not administered to live animals for therapeutic purposes but rather used as a research tool for tissue analysis. Standard protocols for blood smear preparation and staining are followed, with Leishman's stain used at the appropriate concentration.
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| ADME/Pharmacokinetics |
Leishman's stain does not have established pharmacokinetic properties as it is not used as a therapeutic agent. The compound is a histological dye and stain used for research and diagnostic applications. For in vitro use, Leishman's stain is soluble in methanol. The stain is available as a dark-greenish black powder. For storage, the stain should be kept in a cool, dry place and protected from light. The stain is stable under recommended storage conditions.
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| Toxicity/Toxicokinetics |
The toxicity of Leishman's stain is primarily related to its use as a dye and its methanol content. The compound is intended for research and diagnostic use only and is not for human therapeutic use. As a histological dye, Leishman's stain should be handled with appropriate laboratory safety precautions, including the use of gloves and eye protection. The methanol component is flammable and toxic, and appropriate ventilation should be used. The stain may cause irritation to skin, eyes, and respiratory tract upon contact. Inhalation of dust or vapors should be avoided.
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| References | |
| Additional Infomation |
Leishman's stain (CAS 12627-53-1) is a histological stain used for staining blood smears and bone marrow specimens. It is also known as eosin-polychrome methylene blue. Leishman's stain is used to differentiate and identify leukocytes (white blood cells), malaria parasites, and trypanosomes in blood smears. It is a vital dye for staining peripheral blood and bone marrow smears, which appear pale bluish-grey to deep blue when viewed under an oil-immersion lens. The stain is used in hematology, tropical medicine, and cytology. It appears as a dark-greenish black powder and is stored at room temperature. It is intended for research and diagnostic use.
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| CAS # |
12627-53-1
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| Appearance |
Green to dark green solid powder
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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