Wright's stain 

Wright's stain is a composite cell stain that mainly binds to intracellular nucleic acids, proteins and other components through thiazine dyes (such as methylene blue) and eosin. Wright's stain is pH-dependent (optimal pH 6.4-6.7) and achieves cell morphology resolution by differentially staining the cytoplasm and nucleus. Under alkaline conditions, thiazine dyes bind to nucleic acids to form purple, and acidic eosin binds to cytoplasmic proteins to form red, which can form contrasting cell morphological features. Wright's stain can clearly display the fine structures of blood cells and bone marrow cells (such as nuclear chromatin and granules) and quickly evaluate cell morphological abnormalities^[1][2].

1. 骨髓及外周血涂片自动化染色实验：Wright's stain (外周血涂片 12 min，骨髓涂片 20 min) 在 Hematrak 仪器中通过两阶段染色法 (先 Wright's 溶液固定，后 Wright's-缓冲液-Eosin混 合液染色)，使红细胞呈鲜艳橙红色、白细胞核染色质清晰，显著提升细胞形态辨识度，且批间差异小^[1]。
2. 白血病细胞免疫荧光联合检测实验：Wright's stain (常规浓度；40 min) 与上转换纳米颗粒免疫标记结合，在 DLBCL 细胞及临床样本中实现细胞形态 (细胞质/核结构) 与免疫荧光信号的同步观察，辅助鉴别白血病细胞表型，检测限低至 1.54 cell/μL^[2]。

Wright's stain 染色方法
试剂：
(1) 6.3 g Wright's stain 溶于 3.8 L 不含丙酮的甲醇中，陈化 1 个月。
(2) 磷酸盐缓冲液 (pH 6.5) ：将 250 mL 0.067 M?Na2HPO4、250 mL 0.06 M?KH2PO4 和蒸馏水混合至 3 L。
(3) Eosin Y 溶液：12.5 g Eosin Y、575 mL 蒸馏水、1625 mL 95% 甲醇。
步骤固定和初级染色：
将血液/骨髓涂片浸入 Wright's stain 中 3 分钟以固定细胞并开始染色。
缓冲液介导染色：制备 1:5.25 的 Wright's stain 和磷酸盐缓冲液混合物。染色外周血涂片染色 5-8 分钟；骨髓涂片染色 10-15 分钟，以形成细胞核和细胞质对比。
二次 Eosin Y 染色：制备 Eosin Y 溶液、Wright's stain 和缓冲液的混合物 (比例为 1:4:28)。染色涂片 1-2 分钟，以增强红细胞着色 (鲜艳的橙红色) ，同时保留白细胞核细节。
冲洗和干燥：在流动的蒸馏水或 pH 6.3 缓冲液中清洗涂片 2 分钟。在显微镜观察前完全风干。
关键参数最佳 pH：6.5-7.15，可使红细胞和白细胞染色平衡。
总时间：外周血涂片约 12 分钟；骨髓涂片约 20 分钟。

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

68988-92-1

固体

Dark green to black

Room temperature in continental US; may vary elsewhere.

4°C, protect from light

*In solvent : -80°C, 6 months; -20°C, 1 month (protect from light)

• [1]. N Kutaish. Automated staining of bone marrow and peripheral blood by a modified Wright's technic. Am J Clin Pathol. 1982 Mar;77(3):319-20.  [Content Brief]

  [2]. Chen L, et al. A Novel and Rapid Smear Cytomorphology Detection Strategy Based on Upconversion Nanoparticles Immunolabeling Integrated with Wright's Staining for Accurate Diagnosis of Leukemia. Int J Nanomedicine. 2023 Sep 13;18:5213-5224.  [Content Brief]