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Technical Specification: Staining Dyes & Histology Reagents

1. Product Overview

Solarbio staining dyes and histology reagents cover routine H&E staining, special stains for histochemical analysis, and fluorescent dyes for microscopy and flow cytometry. All products are validated on control tissue sections with lot-specific staining patterns documented.

2. Product Range

Dye SKU Application Staining Target Excitation/Emission
Hematoxylin (Harris) G1080 Nuclear counterstain Nuclei (blue-violet) Absorbance-based
Eosin Y (alcoholic) G1100 Cytoplasmic stain Cytoplasm, collagen (pink) Absorbance-based
Hematoxylin (Mayer's) G1085 Nuclear stain Nuclei Absorbance-based
DAPI D1060 Fluorescent nuclear stain DNA 358/461 nm
Propidium Iodide (PI) P8080 DNA content, dead cells DNA 535/615 nm
Hoechst 33342 H1399 Live cell nuclear stain DNA 350/461 nm
Hoechst 33258 H1398 Fixed cell nuclear stain DNA 352/461 nm
Oil Red O O8010 Lipid stain Neutral lipids (red) Absorbance-based
Alcian Blue (pH 2.5) A8170 Mucopolysaccharide stain Acid mucins (blue) Absorbance-based
Congo Red C8090 Amyloid stain Amyloid deposits (red-green birefringence) Absorbance-based
Periodic Acid-Schiff (PAS) Kit G1280 Glycogen/polysaccharide stain Glycoproteins, glycogen (magenta) Absorbance-based
Masson's Trichrome Kit G1340 Connective tissue stain Collagen (blue), muscle (red) Absorbance-based
FITC F8030 Fluorescent labeling Proteins (via conjugation) 495/519 nm
Rhodamine 123 R8030 Mitochondrial membrane potential Active mitochondria 507/529 nm
LysoTracker L8010 Lysosomal staining Acidic organelles 577/590 nm

3.1 H&E Staining (Paraffin Sections)

Step Reagent Time Technical Notes
Deparaffinize Xylene 10 min × 2 Fresh xylene for second wash
Rehydrate 100%→95%→80%→70% ethanol 5 min each Gradual to avoid tissue damage
Nuclear stain Harris hematoxylin 5–8 min Filter before use to remove oxidized surface
Blue in tap water Running tap water 5–10 min Water should be alkaline (pH 8–9) for bluing
Differentiate 1% HCl-EtOH 2–3 s Quick dip, then water rinse
Counterstain Eosin Y 1–2 min Alcoholic eosin preferred for sharper contrast
Dehydrate 95%→100%→xylene 3 min each Complete dehydration critical for permanent mounts
Mount Resinous mounting medium — Use toluene-based; avoid bubble formation

3.2 DAPI Staining

Step Detail Technical Notes
Fix cells 4% paraformaldehyde, 15 min RT Freshly prepared PFA for best nuclear morphology
Wash PBS, 3 × 5 min Gentle agitation
Permeabilize 0.1% Triton X-100, 10 min Adjust for thicker sections: 0.3% Triton, 30 min
Stain DAPI (0.5–1 µg/mL in PBS), 5 min RT, dark Higher concentration for tissue sections (2 µg/mL)
Wash PBS, 3 × 5 min Thorough wash reduces background
Mount Anti-fade mounting medium Use glycerol-based for long-term storage at -20°C

3.3 Oil Red O Staining (Frozen Sections)

Step Detail Time
Section preparation Cryosection at 5–10 µm, air-dry 30 min
Rinse 60% isopropanol 5 min
Stain Oil Red O working solution (0.5% in isopropanol, diluted 3:2 in water) 10–15 min
Rinse 60% isopropanol 5 min (until background clear)
Nuclear counterstain Hematoxylin (Mayer's), optional 1–2 min
Mount Aqueous mounting medium (glycerol-gelatin) —

4. Quality Specifications

Parameter Hematoxylin (Harris) Eosin Y DAPI PI Oil Red O
Purity (HPLC) ≥95% ≥90% ≥98% ≥95% ≥90%
Identity confirmation NMR, UV-Vis NMR, UV-Vis NMR, HPLC NMR, HPLC UV-Vis
Absorbance maximum (λmax) 560 nm (oxidized) 525 nm 358 nm 535 nm 518 nm (in isopropanol)
Emission maximum N/A N/A 461 nm 615 nm N/A
Solubility Water, ethanol Ethanol, water DMSO Water, PBS Isopropanol
pH (1% solution) 2.5–3.5 4.0–5.0 6.0–7.5 5.0–7.0 N/A
Storage RT, light-sensitive RT, light-sensitive -20°C, dark 2–8°C, dark RT, light-sensitive
Shelf life 24 months 24 months 12 months 12 months 24 months

5. Special Stain Kits

Kit SKU Principle Staining Pattern Positive Control Tissue
PAS Stain Kit G1280 Schiff reagent reacts with aldehyde groups Glycogen/mucin: magenta; Nuclei: blue Liver (glycogen), kidney
Masson's Trichrome Kit G1340 Three-dye sequential staining Collagen: blue; Muscle: red; Nuclei: black Skin, heart
Alcian Blue (pH 2.5) A8170 Cationic dye binds to acidic mucins Acid mucins: blue; Background: pale Colon, stomach
Congo Red Stain C8090 Linear dye binds to β-sheet amyloid Amyloid: red-orange; Apple-green birefringence Amyloidotic tissue

6. Troubleshooting Guide

Issue Likely Cause Solution
Hematoxylin fading Oxidized hematoxylin Replace; filter before use
Eosin too intense Overstained or insufficient dehydration Reduce eosin time; ensure complete ethanol steps
DAPI high background Insufficient washing after staining Increase PBS washes to 5 × 5 min
Oil Red O non-specific precipitate Dye crystals formed Filter working solution through 0.45 µm
PAS magenta too weak Periodic acid oxidation insufficient Extend oxidation to 10–15 min at 37°C
Masson's trichrome inconsistent Bouin's fixation inadequate Ensure Bouin's at 56°C for 60 min or overnight at RT

7. Cross-References

For product procurement: solarbio.store