Wright's stain

1. Automated staining of bone marrow and peripheral blood smears: Wright's stain (12 min for peripheral blood smears, 20 min for bone marrow smears) was performed in the Hematrak instrument through a two-stage staining method (first fixation with Wright's solution, then staining with Wright's-buffer-Eosin mixture), which made the red blood cells appear bright orange-red and the white blood cell nuclear chromatin clear, significantly improving the recognition of cell morphology, and with small batch-to-batch differences^[1].
2. Combined immunofluorescence detection of leukemia cells: Wright's stain (conventional concentration; 40 min) was combined with upconversion nanoparticle immunolabeling to achieve simultaneous observation of cell morphology (cytoplasm/nuclear structure) and immunofluorescence signals in DLBCL cells and clinical samples, assisting in the identification of leukemia cell phenotypes, with a detection limit as low as 1.54 cell/μL^[2].

Wright's stain staining method
Reagents:
(1) 6.3 g Wright's stain dissolved in 3.8 L acetone-free methanol, aged for 1 month.
(2) Phosphate buffer (pH 6.5): Mix 250 mL 0.067 M?Na2HPO4, 250 mL 0.06 M?KH2PO4, and distilled water to 3 L.
(3) Eosin Y solution: 12.5 g Eosin Y, 575 mL distilled water, 1625 mL 95% methanol.
Procedure Fixation and Primary Staining:
Immerse blood/bone marrow smears in Wright's stain for 3 minutes to fix cells and initiate staining.
Buffer-mediated staining: Prepare a 1:5.25 mixture of Wright's stain and phosphate buffer. Stain peripheral blood smears for 5-8 minutes; bone marrow smears for 10-15 minutes to provide nuclear and cytoplasmic contrast.
Secondary Eosin Y stain: Prepare a mixture of Eosin Y solution, Wright's stain, and buffer (1:4:28 ratio). Stain smears for 1-2 minutes to enhance red blood cell staining (bright orange-red) while preserving white blood cell nuclear detail.
Rinse and dry: Wash smears in running distilled water or pH 6.3 buffer for 2 minutes. Air dry completely before microscopic observation.
Key parameters Optimal pH: 6.5-7.15, which allows for balanced red and white blood cell staining.
Total time: approximately 12 minutes for peripheral blood smears; approximately 20 minutes for bone marrow smears.

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.