Thionin Staining Solution (2%) - BioReagent,Biological Stain,for microscopy,2% , CAS No.78338-22-4

CAS: 78338-22-4 Cat. No.: S1508553 분자식: C₁₂H₉N₃S·C₂H₄O₂ 분자량: 287.34 EC 번호: 676-854-9 PubChem CID: 2724414

등급 & 순도 BioReagent, Biological Stain, for Microscopy, 2%

Synonyms

Thionin Blue Staining Solution | Thionine Staining Solution

Storage

Protected from light,Room temperature

Shipped In

Normal

Application

Cell Staining

개요

Many dyes exhibit varying degrees of affinity for DNA and can therefore be used to stain chromosomes. For routine staining of chromosomes, good results are generally obtained with Giemsa, orecin, Feulgen, carbol fuchsin, etc. Thionine can be used for chromosome staining in samples such as oral mucosa, urine, amniotic fluid, chorionic villus cells, and cultured cells, and is particularly suitable for staining sex chromatin, which is often difficult to stain. Additionally, thionine can be used for staining structures such as Nissl bodies and mast cells.

Thionin Staining Solution (2%) is primarily composed of phosphate and thionine. When staining Nissl bodies, they appear dark blue; however, when staining mast cells, they appear purplish‑red. This reagent is intended for research use only and is not suitable for clinical diagnosis or other purposes.

Instructions for Use (For Reference Only)

1. Staining of Nissl Bodies

• Fixation: Use neutral formalin fixative or other appropriate fixatives.

• Paraffin sections 6–8 μm thick; routine deparaffinization with xylene or dewaxing clearing solution to distilled water.

• Immerse the slide specimen in Thionine Staining Solution (2%) for 10–15 minutes; rinse briefly with distilled water.

• Differentiate rapidly with 95% ethanol.

• Dehydrate with absolute ethanol, clear with xylene or dewaxing clearing solution, and mount with neutral balsam.

2. Staining of Chromatin

• Place the slide specimen in 1 mol/L HCl and incubate at 37°C for 20 minutes.

• Rinse thoroughly with distilled water and air‑dry.

• Immerse the slide specimen in Thionine Staining Solution (2%) for 10–15 minutes.

• Rinse with distilled water and air‑dry.

• Under low magnification, search for evenly dispersed cell populations, then carefully observe under oil immersion.

Staining Results

1. Nissl bodies appear dark blue; nuclei appear light blue.

2. Chromatin appears purplish‑red; other components appear blue.

Notes

1. Tissue intended for Nissl body staining should be as fresh as possible and fixed promptly; otherwise, Nissl bodies may dissolve and stain poorly.

2. Nissl body specimens should be stored protected from light, as they are prone to fading.

3. During centrifugation to obtain cell pellets, for certain cell types, if the pellet is insufficient, centrifugation force or time may be increased appropriately.

4. This staining solution is more suitable for Nissl body staining; although it can be used for chromatin staining, it is not recommended.

5. For your safety and health, please wear a lab coat and disposable gloves during operation.

6. Use the reagents as soon as possible after opening to avoid affecting subsequent experimental results.

사양

동의어

Thionin Blue Staining Solution | Thionine Staining Solution

사양 및 순도

BioReagent,Biological Stain,for microscopy,2%

안정성 및 스토리지

Store at room temperature long term (12 months). Store in the dark.

보관 조건

Protected from light,Room temperature

배송

Normal

등급

Biological Stain, BioReagent, 현미경용

Product Line

생물 제품

이름과 식별자

이성체 SMILES

CC(=O)[O-].C1=CC2=C(C=C1N)SC3=CC(=[NH2+])C=CC3=N2

PubChem CID

2724414

분자량

287.34

화학 및 물리적 특성

• 감도: Light-sensitive

Application Protocols

The following protocol is provided as general literature guidance for Nissl staining with a thionin 2% solution. Adjust to your specimen, fixation, and instrumentation. This is not an item-specific validated protocol.

Example protocol (paraffin sections):

1. Deparaffinize slides in xylene (2×, 5 min each) and rehydrate through graded ethanols to water.
2. Optional: Pre-treat in acetate buffer (pH ~4.5–5.0) for 1–3 min to improve selectivity (if desired by your method).
3. Stain in thionin staining solution (2%) for 1–5 min at room temperature. Begin with 1 min for thin sections.
4. Rinse briefly in distilled water.
5. Differentiate in 95% ethanol, optionally containing 0.5–1% acetic acid, with monitoring under a dissecting microscope until background is acceptably low.
6. Rinse quickly in 100% ethanol to stop differentiation; proceed to 2× changes of 100% ethanol to complete dehydration.
7. Clear in xylene or substitute (2×) and mount with a resinous medium.

Semi-thin resin sections (literature):

• Short stain (seconds to 1 min), immediate differentiation in ethanol, rapid dehydration/clearing.

Controls and QA:

• Include an unstained control and a known positive control tissue.
• Record times, temperatures, and bath age; replace staining bath when color strength diminishes or contamination is evident.

Biological Roles

This product is a staining solution; it does not possess a biological role in vivo. The following notes describe interactions relevant to microscopy (literature):

• Electrostatic interactions: Thionin is a cationic chromophore that binds polyanionic biomolecules (nucleic acids, RNA-rich ribosomes/Nissl substance) via electrostatic attraction and stacking interactions, leading to strong nuclear and nucleolar coloration.
• Metachromasia and matrix effects: In matrices rich in sulfated glycosaminoglycans (e.g., cartilage), cationic dyes can exhibit metachromasia (shift in color due to dye–dye and dye–polymer interactions), though thionin is less classically metachromatic than toluidine blue O. Extent depends on concentration, fixation, and pH.
• Photochemistry: Phenothiazinium dyes can undergo photoinduced redox processes and photobleaching. During microscopy, minimize high-intensity light exposure to preserve stain intensity.
• Compatibility with fixation: Formaldehyde and glutaraldehyde fixatives generally preserve nucleic acids and ribosomes, supporting robust staining. Over-fixation or harsh decalcification may diminish intensity.
• Counterstaining: Thionin can serve as a primary nuclear stain or as a counterstain in multi-stain protocols, provided spectral contrast is maintained with other chromogens or fluorophores.

These behaviors influence staining selectivity and intensity but do not imply any physiological or therapeutic function. Optimize conditions empirically for your specimen type.

Buffer Applications

Not typically applicable. This product is a ready-to-use staining solution rather than a buffering agent.

Practical notes (literature):

• If you require pH-controlled staining, use separate buffer baths (e.g., acetate buffers in the mildly acidic range) before or after exposure to the dye to modulate selectivity and background.
• Rinse steps in deionized water or buffer can help remove non-specifically bound dye and standardize outcomes across lots.

For buffer compositions, pH ranges, and recipes, consult validated histology protocols appropriate to your tissue type and fixation.

Green Alternatives

While this item is a ready-to-use dye solution rather than an organic solvent or reagent, greener practice considerations can still improve environmental and operator safety performance.

• Waste minimization: Use small-volume Coplin jars or slide racks to reduce bath volumes. Reuse working baths judiciously with filtration, recognizing that dye depletion and carryover can shift outcomes.
• Safer clearing and dehydration: Consider xylene substitutes with improved safety profiles for post-stain clearing. Ethanol graded series remains standard; isopropanol may substitute in some workflows.
• Alternative dyes (literature comparison; selection depends on target and matrix):

Comparison (literature; not item specifications)

• Thionin (this product): Strong nuclear/Nissl stain; vivid violet; good for semi-thin resin sections; may photofade; cationic.
• Cresyl violet: Widely used Nissl stain; often robust with clear nucleolar contrast; similar handling; may be less prone to over-staining in some matrices.
• Toluidine blue O: Rapid, metachromatic staining for semi-thin sections; excellent for resin-embedded tissues; high sensitivity to acidic components; blue tone may be preferable.
• Methylene blue/azure mixtures: Effective for quick screening; different hue/contrast; sometimes lower hazard classifications.

Trade-offs:

• Hue and metachromasia differ across dyes, affecting feature recognition.
• Some alternatives may present lower environmental persistence or toxicity, but performance on your specimen type should guide selection.

Always consult SDS and institutional green-chemistry guidance when selecting clearing agents and mounting media.

Pharmaceutical Uses

Not applicable. This is a laboratory staining solution for microscopy and is labeled For research use only (Product Data). It is not formulated, tested, or intended for pharmaceutical manufacturing or as an excipient.

Process-related note (general): If similar dyes are used in diagnostic manufacturing or device QC, their use is governed by specific quality standards and validations that are beyond the scope of this research-grade reagent.

Physical Properties

Item-specific physical specifications for this ready-to-use solution are not provided. Where relevant, general/literature properties of thionin and its aqueous solutions are noted and clearly labeled.

Item-specific (from Product Data):

• Physical state/appearance: Not specified for this item; refer to CoA/Spec Sheet. (Typical: violet aqueous solution; literature)
• Concentration: 2% (Product name)
• Storage: Room temperature, protected from light (Product Data)

General/literature values (thionin chromophore/aqueous solutions; not item specifications):

• Solubility: Highly soluble in water; miscible with common aqueous buffers. Limited solubility in nonpolar solvents due to cationic nature.
• UV/Vis absorption: Strong visible absorption; reported λmax commonly near ~596–600 nm in water with secondary bands in the UV. Exact spectrum depends on pH/ionic strength.
• pH dependence: Protonation state and spectral properties vary with pH; enhanced binding to polyanions (e.g., nucleic acids) in neutral to slightly acidic media.
• Physical hazards: Dye readily stains skin and labware; may adsorb to glass and plastics.

Not specified for this item; refer to CoA/Spec Sheet:

• Density, refractive index, pH range, conductivity, viscosity, freezing point, and stabilizers (if any).

Quality and Grades

• Grade/Purity (Product Data): BioReagent, Biological Stain, for Microscopy.

What this implies:

• BioReagent: Suitable for common life-science workflows; manufactured to support reproducible staining in histology/cytology and related microscopy workflows.
• Biological Stain (for Microscopy): Formulated to deliver consistent chromatic performance and contrast on biological specimens. Emphasis is on dye performance (color strength, contrast, minimal background) rather than chromatographic purity metrics.

Item-specific notes:

• This is a ready-to-use 2% solution. The exact solvent system, pH, and any stabilizers are Not specified for this item; refer to CoA/Spec Sheet.
• Appearance, absorbance specification, particulate/filtration status, and lot-to-lot tolerances are Not specified for this item; refer to CoA/Spec Sheet.

Practical guidance for users:

• For quantitative imaging, consider recording baseline absorbance spectra of each lot to normalize exposure times.
• If background staining is observed in your matrix, pre-filter through 0.22 µm PVDF/nylon (if compatible) immediately before use; verify compatibility on a small aliquot first.
• For techniques sensitive to UV background (e.g., fluorescence counterstaining), ensure complete washing after thionin application to minimize bleed-through in overlapping channels.

Documentation:

• For regulated workflows, retain the CoA and SDS for your specific lot and verify that the stated grade aligns with your method requirements.

Reaction and Applications

This product is intended for microscopy staining, not for chemical synthesis. The following are application-focused uses in biological sample preparation (literature/good practice):

• Nissl staining of nervous tissue: Thionin, a cationic phenothiazinium dye, binds ribosomal RNA and nucleic acids to visualize Nissl substance in neurons, enabling cytoarchitectonic analysis in brain and spinal cord sections.
• General nuclear/cytoplasmic contrast: Provides strong nuclear staining in many tissues due to affinity for polyanionic biomacromolecules (DNA/RNA), assisting in cell counting and morphology assessment.
• Resin-embedded semi-thin sections: Frequently used on epoxy- or acrylic-embedded sections (e.g., 0.5–1 µm) for rapid screening under brightfield.
• Botanical and microbiological preparations: Binds acidic polysaccharides and nucleic acids, offering contrast in certain plant tissues and smears (protocol-dependent).

Practical tips (literature):

• Pre-treatment: Thorough deparaffinization and hydration (for FFPE) or rehydration (for cryosections) improves uniform dye access.
• Control of selectivity: Slightly acidic baths can enhance nucleolar/Nissl selectivity; differentiation in 95% ethanol (optionally with acetic acid) reduces background.
• Timing: Staining times vary widely (from ~30 s to several minutes) depending on section thickness, fixation, and matrix.
• Compatibility: After staining and differentiation, standard dehydration and clearing steps precede resinous mounting media. Avoid prolonged light exposure during and after staining to limit fading.

Note: Optimize parameters empirically for your specimen and microscope settings.

Reaction Conditions

Not applicable to chemical synthesis. For staining workflows, typical “conditions” correspond to histology parameters (literature guidance only):

• Section preparation: Deparaffinize FFPE sections and hydrate to water; for resin or cryosections, ensure appropriate rehydration.
• Staining time: Commonly 0.5–10 minutes depending on section thickness, fixation, and desired intensity. Start with short exposures and increase as needed.
• Differentiation: Brief dips to minutes in 95% ethanol (optionally with 0.5–1% acetic acid) to reduce background and enhance nuclear contrast.
• Rinsing: Water rinse between steps to halt staining and remove unbound dye.
• Dehydration and clearing: Graded ethanols to absolute, then clearing agent prior to resinous mounting media.
• Temperature: Ambient.

These are general literature parameters, not specifications for this item. Optimize empirically for your specimen and imaging setup.

Safety and Handling

Authoritative safety information is provided in the SDS for this catalog item. The following guidance is general, based on typical handling of cationic biological stains and the Product Data.

Product Data specifics:

• Storage conditions: Protect from light; store at room temperature.
• Shipping: Normal (ambient).

GHS classification, signal word, pictograms, and H-statements: Not specified for this item; refer to the SDS.

General laboratory precautions (literature/best practice for biological stains):

• PPE: Wear appropriate gloves, lab coat, and eye protection. Thionin strongly stains skin, fabrics, and porous surfaces—use dedicated staining vessels and secondary containment.
• Ventilation: Use in a well-ventilated area; avoid aerosol generation and inhalation of mists.
• Incompatibilities: Avoid strong oxidizing or reducing agents that can degrade the dye; avoid strong bases/acids that may alter staining performance.
• Light sensitivity: Protect from light to minimize photobleaching and color shift.
• Spill/cleanup: Absorb small spills with inert material; decontaminate surfaces with dilute bleach or appropriate detergent, followed by thorough rinsing. Dispose of dye-contaminated waste according to institutional and local regulations.
• First aid (overview; consult SDS): In case of skin contact, wash with soap and water. For eye exposure, rinse cautiously with water for several minutes and seek medical attention. If ingested, rinse mouth; do not induce vomiting; seek medical advice.

For research use only (Product Data).

Solvent Selection

This product is supplied as a ready-to-use 2% staining solution; separate solvent selection is generally not required.

Relevant solvent characteristics (general/literature for thionin solutions):

• Primary medium: Aqueous (typical). The cationic dye exhibits high water solubility and is compatible with common buffer salts.
• Miscibility and processing: After staining, sections are often differentiated and dehydrated in graded ethanols and cleared in xylenes or substitutes prior to mounting (literature practice). Verify compatibility of your workflow with the supplied solution.

When would alternative media be considered?

• Alcoholic thionin solutions may be favored for rapid penetration in resin-embedded or highly hydrophobic matrices (literature). If your protocol requires alcoholic formulations, confirm whether this item’s composition is suitable; otherwise prepare a validated alternative.
• pH-adjusted aqueous buffers (e.g., acetate) can modulate selectivity for nucleic acids vs. background (literature). If pH control is critical, assess the behavior of this solution versus a freshly prepared, pH-defined bath.

Non-applicable details:

• Traditional “solvent selection” for synthetic reactions does not apply. For staining workflows, focus instead on pre- and post-stain steps (hydration, differentiation, dehydration, clearing) that determine outcome and contrast.

Storage and Reconstitution

Item-specific (Product Data):

• Storage conditions: Store at room temperature; protect from light to minimize photodegradation and color shift.
• Shipping: Normal (ambient).
• Reconstitution: Not applicable. This is a ready-to-use 2% staining solution.

Good laboratory practice:

• Keep the container tightly closed when not in use to limit evaporation and contamination.
• Minimize exposure to strong light sources; use amber bottles or store in the original light-protective container.
• Do not freeze unless explicitly supported by the CoA/Spec Sheet, as freeze–thaw can precipitate components or alter performance.
• If particulates are observed, pre-filter a working aliquot through a 0.22–0.45 µm compatible membrane; do not filter the entire stock unless validated.
• Assign in-lab open/expiry dating based on your QC criteria. If the CoA provides a retest date, follow that guidance.

Unspecified for this item; refer to CoA/Spec Sheet:

• Exact shelf life, permissible temperature excursions, and any stabilizers/preservatives.

For research use only (Product Data).

Structure and Identity

This product is a ready-to-use 2% Thionin Staining Solution formulated for microscopy.

• SKU: S1508553; Product name: Thionin Staining Solution (2%)
• CAS: 78338-22-4 (assigned to thionin staining solutions)
• Grade/Use: BioReagent, Biological Stain, for Microscopy (Product Data)
• Active component: thionin dye (phenothiazinium cationic dye; literature)
• Molecular formula: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight: Not specified for this item; refer to CoA/Spec Sheet.
• SMILES: Not specified for this item; refer to CoA/Spec Sheet.
• InChIKey: Not specified for this item; refer to CoA/Spec Sheet.

Structural features (literature, for the thionin chromophore):

• Phenothiazin-5-ium heteroaromatic core with two ring nitrogens and one sulfur in the tricyclic framework.
• Typically bears amino substituents; overall cationic chromophore balanced by an anion (e.g., chloride/acetate in many commercial preparations). Exact counterion for this item is not specified.
• Conjugated, planar aromatic system responsible for intense violet coloration and strong absorption in the visible range.

2D description in words (literature):

• Three fused rings (two benzenoid rings flanking a central 1,4-thiazinium-like ring), para- and meta- amino substituents on the outer rings, yielding a delocalized cation over the tricyclic system. As sold here, the dye is present as a 2% solution; exact excipients/solvent composition are not specified for this item.

Synthetic Utility

Not applicable. This product is a prepared 2% staining solution intended for microscopy workflows, not a reagent for synthetic organic chemistry.

If you require thionin as a chemical reagent (e.g., as a photoredox photosensitizer in electron-transfer studies; literature), procure a solid dye with defined counterion and purity specifications suitable for reaction use rather than a ready-to-use staining solution.

Target Specificity

Not applicable. This product is not an antibody or affinity reagent. It is a general cationic biological stain with broad affinity for polyanionic biomacromolecules (e.g., nucleic acids; literature). No antigen/epitope specificity applies.