Neutral Red Staining Solution (Vacuolar System Specific) - BioReagent,for microscopy,Biological Stain , CAS No.553-24-2

CAS: 553-24-2 Cat. No.: N1513165 Molecular Weight: 288.78 EC Number: 209-035-8

GRADE & PURITY BioReagent, Biological Stain, for Microscopy

Synonyms

Neutral Red Staining Solution (Specific for Vacuolar System Staining)

Storage

Room temperature

Shipped In

Normal

Application

Cell Staining

Overview

  Neutral red is a weakly basic pH indicator and a commonly used vital stain. Its color change range is pH 6.4–8.0 (red to yellow). Its molecular formula is C₁₅H₁₆N₄·HCl, and its molecular weight is 288.78. As a reagent for specific staining of the vacuolar system, its staining effect is closely related to cell viability, intracellular environment, and cell type. The specific characteristics and staining principles are as follows:

  For plant cells, under neutral or slightly alkaline conditions, living cells can absorb large amounts of neutral red and excrete it into the vacuole. Since the vacuole of plant cells typically exhibits an acidic reaction, the neutral red that enters the vacuole dissociates into a large number of cations, resulting in a cherry-red color. During this process, the protoplasm and cell wall generally remain unstained. In dead cells, due to the denaturation and coagulation of the protoplasm, the cell sap cannot be maintained within the vacuole, and no vacuolar staining occurs after neutral red staining. Instead, the cations of neutral red bind to the negatively charged protoplasm and nucleus, staining both.

  For animal cells, the vacuolar system consists of vesicles surrounded by a single membrane, including lysosomes, the Golgi complex, the endoplasmic reticulum, phagosomes, transport vesicles, etc. The vacuolar system is most developed in chondrocytes, and neutral red can specifically stain it.

  The Neutral Red Staining Solution (Vacuolar System Specific) is neutral in pH. As a specific vital stain for the vacuolar system in tissues or cells, when staining living cells, it only stains the vacuolar system red, while the cytoplasm and nucleus remain unstained. This allows for precise and specific observation of the vacuolar system.

Product Component List

N1513165	Component	100 mL	Storage
N1513165A	Neutral Red Staining Solution (Vacuolar System Specific)	100 mL	RT.
N1513165B	Ringer Buffer	250 mL	RT.

Procedure (For Reference Only)

1. Sacrifice a toad or other animal by severing the spinal cord. Secure it on a wax dish, open the abdominal cavity, and obtain a small piece from the thinnest part of the xiphoid cartilage. Place it on a clean glass slide.

2. Add the Neutral Red Staining Solution (for Vacuolar System), ensuring the sample is completely covered by the stain. Stain for 15–20 minutes.

3. Use absorbent filter paper to remove excess stain from around the edges of the slide.

4. Add Ringer Buffer, cover with a coverslip, and use filter paper to absorb excess liquid from the side of the coverslip.

5. Immediately after completion, proceed to microscopic observation or photography.

Staining Results

  Under the microscope, chondrocytes appear oval. The intracellular vacuolar system consists of vesicles of varying sizes, exhibiting a rose-red color. The cytoplasm and nucleus remain unstained, allowing clear distinction of the vacuolar system structures.

Notes

1. Long-term storage of the Neutral Red Staining Solution may produce a small amount of precipitate, which generally does not affect experimental use.

2. When using this reagent for the first time, it is recommended to perform a pilot experiment with 1–2 samples to confirm the staining effect and compatibility before proceeding with large-scale operations.

3. During staining, the staining time can be adjusted flexibly according to the actual staining results and experimental requirements.

4. This experiment involves vital staining. During sample collection, ensure accuracy and speed, and maintain the vital state of the sample throughout the process to avoid affecting the staining results.

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

6. This product is intended for research use only and is strictly prohibited for any other purposes.

Specifications

Synonyms

Neutral Red Staining Solution (Specific for Vacuolar System Staining)

Specifications & Purity

BioReagent, for microscopy, Biological Stain

Stability And Storage

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

Storage

Room temperature

Shipped In

Normal

Grade

Biological Stain, BioReagent, for Microscopy

Names and Identifiers

• Isomeric SMILES: CC1=CC2=NC3=C(C=C(C=C3)N(C)C)N=C2C=C1N.Cl
• Molecular Weight: 288.78
• Reaxy-Rn: 3918740
• Reaxys-RN_link_address: https://www.reaxys.com/reaxys/secured/hopinto.do?context=S&query=IDE.XRN=3918740&ln=

Chemical and Physical Properties

• Melt Point(°C): 290°C

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64. Jianliang Xiao, Weiyang Lv, Zhou Xie, Yeqiang Tan, Yihu Song, Qiang Zheng.  (2016)  Environmentally friendly reduced graphene oxide as a broad-spectrum adsorbent for anionic and cationic dyes via π–π interactions.  Journal of Materials Chemistry A,  4  (31): (12126-12135).  [PMID:] [10.1039/C6TA04119A]

65. Lili Ren, Yingchun Cai, Limin Ren, Hong Yang.  (2016)  Preparation of Modified Beeswax and Its Influence on the Surface Properties of Compressed Poplar Wood.  Materials,  9  (4): (230).  [PMID:28773357] [10.3390/ma9040230]

66. Hong Niu, Dan Song, Haibo Mu, Wuxia Zhang, Feifei Sun, Jinyou Duan.  (2016)  Investigation of three lignin complexes with antioxidant and immunological capacities from Inonotus obliquus.  INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,      [PMID:26845476] [10.1016/j.ijbiomac.2016.01.111]

67. Yan Zhao, Weijia Cao, Zhen Wang, Bowen Zhang, Kequan Chen, Pingkai Ouyang.  (2015)  Enhanced succinic acid production from corncob hydrolysate by microbial electrolysis cells.  BIORESOURCE TECHNOLOGY,      [PMID:26708482] [10.1016/j.biortech.2015.12.002]

68. Linlin Wang, Yizhong Shen, Shaopu Liu, Jidong Yang, Wanjun Liang, Dan Li, Youqiu He.  (2014)  Fluorescent reversible regulation based on the interactions of topotecan hydrochloride, neutral red and quantum dots.  SPECTROCHIMICA ACTA PART A-MOLECULAR AND BIOMOLECULAR SPECTROSCOPY,      [PMID:25459722] [10.1016/j.saa.2014.10.054]

69. Sui Wang, Jing Wei, Shasha Lv, Zhiyong Guo, Feng Jiang.  (2013)  Removal of Organic Dyes in Environmental Water onto Magnetic-Sulfonic Graphene Nanocomposite.  CLEAN-Soil Air Water,  41  (10): (992-1001).  [PMID:] [10.1002/clen.201200460]

70. Tingting Wang, Fanghui Ma, Feng Huang, Aomei Zhang, Minghui Yang, Xiaoqing Li, Xiang Chen.  (2024)  A background correctable fluorescence sensing strategy for adenosine triphosphate evaluation based on aptamer configuration alteration.  SENSORS AND ACTUATORS B-CHEMICAL,      [PMID:] [10.1016/j.snb.2024.135306]

71. Xiaodong Zhai, Yuhong Xue, Wenjun Song, Yue Sun, Tingting Shen, Xinai Zhang, Yanxiao Li, Fuyuan Ding, Di Zhang, Chenguang Zhou, Muhammad Arslan, Haroon E. Tahir, Zhihua Li, Jiyong Shi, Xiaowei Huang, Xiaobo Zou.  (2024)  A ratiometric fluorescent electrospun film with high amine sensitivity and stability for visual monitoring of livestock meat freshness.  Food Chemistry-X,      [PMID:39290751] [10.1016/j.fochx.2024.101801]

72. Qiuhui Qian, Qian Pu, Xin Li, XingCheng Liu, Anyu Ni, Xiaowen Han, Zejun Wang, Xuedong Wang, Jin Yan, Huili Wang.  (2024)  Acute/chronic triclosan exposure induces downregulation of m6A-RNA methylation modification via mettl3 suppression and elicits developmental and immune toxicity to zebrafish.  CHEMOSPHERE,      [PMID:38342143] [10.1016/j.chemosphere.2024.141395]

73. Huilin Chen, Ran Xu, Enyu Xu, Yan Chen, Chunyu Niu, Yin Chen.  (2024)  Construction and performance evaluation of polyguluronic acid polysaccharides-based drug delivery systems.  COLLOIDS AND SURFACES B-BIOINTERFACES,      [PMID:39029246] [10.1016/j.colsurfb.2024.114083]

74. Feng Li, Kairen Zhao, Yan Jin, Baoxin Li.  (2024)  Efficient degradation of organic pollutants without any external assistance over a wide pH range using carbon vacancy-modified Fe–N–C catalysts.  Nanoscale,  16  (48): (22273-22282).  [PMID:39534988] [10.1039/D4NR03755C]

75. Wang Cong, Chang Dongdong, Zhang Qi, Yu Zhisheng.  (2024)  Enhanced bioethanol production by evolved Escherichia coli LGE2-H in a microbial electrolysis cell system.  Bioresources and Bioprocessing,  11  (1): (1-10).  [PMID:38647898] [10.1186/s40643-023-00717-5]

76. Li Mengke, Cao Panpan, Dong Wenwen, Zhao Jun, Li Dongsheng.  (2024)  Fast and Efficient Removal of Anionic Dyes from Water by Hierarchical Porous MIL-101(Cr).  Journal of Inorganic and Organometallic Polymers and Materials,      [PMID:] [10.1007/s10904-024-03252-8]

77. Linglian Kong, Peng Sun, Xue Pan, Chuanpi Xiao, Bochen Song, Zhigang Song.  (2024)  Glycerol monolaurate regulates apoptosis and inflammation by suppressing lipopolysaccharide-induced ROS production and NF-κB activation in avian macrophages.  POULTRY SCIENCE,      [PMID:38851181] [10.1016/j.psj.2024.103870]

78. Wang Yi, Hu Yaozhong, Niu Zhenhua, Zhang Xuejiao, Fan Dancai, Ji Xuemeng, Lv Huan, Wang Shuo, Zhao Yanrong.  (2024)  Immunomodulation of nutritional formula containing epigallocatechin-3-gallate, ginseng extract, and polydextrose on inflammation and macrophage polarization.  Frontiers in Nutrition,      [PMID:38445210] [10.3389/fnut.2024.1370608]

79. Yunjin Yao, Zhongming Tao, Hongwei Hu, Lijie Zhang, Ziwei Ma, Yaoyao Wang, Shiyang Lin, Shaobin Wang.  (2024)  In situ growth of iron incorporated Ni3S2 nanosheet on nickel foam in mediating electron transfer to peroxymonosulfate for pollutant abatement.  Journal of Environmental Sciences,      [PMID:39306441] [10.1016/j.jes.2024.03.054]

80. Lei Ge, Jiaqi Li, Gaojing Du, Kai Sheng, Jingwei Hou, Binyu Zhou, Yatao Zhang, Bart Van der Bruggen, Junyong Zhu.  (2025)  Microporous Polyacylhydrazone Membranes with 3D Interconnected Nanochannels for Accurate Molecular Sieving.  ADVANCED FUNCTIONAL MATERIALS,      [PMID:] [10.1002/adfm.202504997]

81. Xiaoyu Yang, Hongdan Wu, Zhihui Zhou, Yehao Zhao, Yuedong Li.  (2024)  Modification of a MoS2 composite membrane using nano-silica for high-efficiency dye separation.  NEW JOURNAL OF CHEMISTRY,  48  (16): (7439-7448).  [PMID:] [10.1039/D3NJ05932D]

82. Yunjin Yao, Wei Wang, Hongwei Hu, Lijie Zhang, Ziwei Ma, Shaobin Wang.  (2024)  Nickel–manganese sulfide on nickel foam toward highly efficient and stable potassium periodate activation for organic pollutant degradation.  CHEMICAL ENGINEERING SCIENCE,      [PMID:] [10.1016/j.ces.2024.120276]

83. Dandan Xie, Xiaochuan Yang, Hang Wang, Qiuxia Fu, Fayun Wei, Nuo Liu, Hailou Wang, Guangyu Zhang, Jiamu Dai, Chunhong Zhu, Wei Zhang.  (2024)  Non-destructive strategy to extract sustainable helix and high-strength Musa core fibers for rapid water conduction and evaporation.  INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,      [PMID:38734352] [10.1016/j.ijbiomac.2024.132276]

84. Jianming Wu, Shuai Chen, Qinjie Xu, Qingkai Pang, Peng Li, Yulin Li.  (2024)  Programable sewage-cleaning technology: Regenerating chitosan biofilms with anti-bacterial capacity via self-purification of water pollutants.  INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES,      [PMID:38754677] [10.1016/j.ijbiomac.2024.132355]

85. Qian Lei, Yaoyu Zhang, Tongtong Yang, Lei Chen, Xinxin Pei, Yujie Zhang, Tengfei Ma, Yuzhu Xie, Yi Wang, Hongxia Li, Cheng Cao, Buchang Zhang, Changzhi Xu.  (2024)  Supplementation of dietary heptadecanoic acid enhances anti-Listeria monocytogenes response in macrophages.  Journal of Functional Foods,      [PMID:] [10.1016/j.jff.2024.106359]

86. Shizong Wang, Jianlong Wang.  (2024)  The correlation between electron exchange capacity of Fenton-like heterogeneous catalyst and catalytic activity.  CHEMOSPHERE,      [PMID:38494002] [10.1016/j.chemosphere.2024.141587]

87. Li Haiyang, Li Xiaran, Quan Fengyu, Chen Zhaojun, Xia Yanzhi, Xiong Zhong.  (2024)  Wide-Spectrum Dye Adsorption Performance of Alginic Acid Carbon Aerogel.  WATER AIR AND SOIL POLLUTION,  235  (6): (1-12).  [PMID:] [10.1007/s11270-024-07181-7]

88. Xingdi Zhao, Pengfei Bian, Liu Wang, Xinyu Li, Jingxin Zhou, Yuqing Qiao, Mingli Wang, Tifeng Jiao.  (2025)  SERS and photoelectric conversion properties of cholesterol derivatives composite Langmuir-Blodgett films with dyes.  COLLOIDS AND SURFACES A-PHYSICOCHEMICAL AND ENGINEERING ASPECTS,      [PMID:] [10.1016/j.colsurfa.2025.137741]

89. Tianyi Hu, Qun Liu, Zeng Xu, Zhengjiang Zhu, Wuyin Weng, Yucang Zhang.  (2025)  Antibacterial starch-cellulose-based films with pH-responsive colorimetric properties for real-time food freshness monitoring.  CHEMICAL ENGINEERING JOURNAL,      [PMID:] [10.1016/j.cej.2025.166723]

90. Xinxin Tong, Zihan Li, Xiaoqing Li, Zongyang Tan, Xiaoyu Liu, Xujia Zhao, Mingyang Zhou, Yongqian Lei, Pengran Guo, Jian Liu, Shenghong Yang.  (2025)  Solid-phase engineered red-emissive carbon dots for multi-scenario uranyl ion tracking and on-demand pH-triggered antibacterial.  JOURNAL OF HAZARDOUS MATERIALS,      [PMID:41016207] [10.1016/j.jhazmat.2025.139959]

91. Xiaoyu Yang, Shunbi Xie, Weifen Wu, Jingwen Liu, Manjun Liao, Runzi Zhang, Yao Liu, Mengjun Wang, Yi He.  (2025)  Core-shell structure and twin-cone morphology of Au@Ag@HOF with efficient adsorption capability and high SERS enhancement efficiency for sensitive detection of bisphenol compounds in food.  FOOD CHEMISTRY,      [PMID:41027209] [10.1016/j.foodchem.2025.146522]

92. Hai-Chuan Yu, Meng-Yao Chen, Shuang-Ling Zhang, Fang-Chao Tong, Fan-Ping Wang, Lin Wang, Jia-Long Zhang, Jiao-Jie Guo, Bing-Yang Zhang, Jia-Qi Tian, Jiao Wu.  (2025)  LPS disrupts erythroid-myeloid balance in zebrafish via Jak2/Stat3-Hif1a signaling.  FISH & SHELLFISH IMMUNOLOGY,      [PMID:40935171] [10.1016/j.fsi.2025.110874]

93. Yunshuo Chen, Yachan Gong, Dan Li, Weiwei Feng, Yao Chen, Ting Zhao, Liuqing Yang, Guanghua Mao, Xiangyang Wu.  (2025)  Dinotefuran exposure induces immunotoxicity in zebrafish embryos.  COMPARATIVE BIOCHEMISTRY AND PHYSIOLOGY C-TOXICOLOGY & PHARMACOLOGY,      [PMID:40222697] [10.1016/j.cbpc.2025.110206]

94. Xincong Su, Li Kai, Xiaowen Han, Rongzhi Wang, Xiao Yang, Xuedong Wang, Jin Yan, Qiuhui Qian, Zejun Wang, Huili Wang.  (2025)  Equipotent bisphenol S and bisphenol F with widely differing modes of action exhibit additive effects in immunotoxicity: insights based on intrinsic immunity, apoptosis and regeneration, and oxidative stress.  SCIENCE OF THE TOTAL ENVIRONMENT,      [PMID:40239502] [10.1016/j.scitotenv.2025.179405]

95. Tianyi Hu, Qun Liu, Shoumei Wan, Liangting Deng, Xu Zeng, Wuyin Weng, Yucang Zhang.  (2025)  Development of a multifunctional starch/cellulose-based film with superior mechanical strength, antibacterial activity and visual food freshness indication.  Materials Today Communications,      [PMID:] [10.1016/j.mtcomm.2025.112538]

96. Hongjuan Bai, Dan Liu, Gang Wang, Yuanyuan Feng, Xuanyan Zhu, Huiling Lu, Pengzhan Guo, Yifei Ding, Shuai Yang, Junhang Chen.  (2025)  A novel sulfonated polybenzothiazole-based adsorbent for simultaneous capture of organic dyes from aqueous solution.  SEPARATION SCIENCE AND TECHNOLOGY,      [PMID:] [10.1080/01496395.2025.2502749]

97. DongXue Ma, YuXin Zhang, Jia Zhang, Jun Shi, ShanHu Gao, Fei Long, Xin Wang, XingYu Pu, Jiayao Sun, Shuang Liang, Richard D. Cannon, Silas Villas-Boas, Ting-Li Han.  (2025)  Outer membrane vesicles derived from probiotic Escherichia coli Nissle 1917 promote metabolic remodeling and M1 polarization of RAW264.7 macrophages.  Frontiers in Immunology,      [PMID:40510339] [10.3389/fimmu.2025.1501174]

98. Dan Li, Xiaolong Li, Zijia Huang, Tengteng Liu, Mao Li, Xiaoming Chen, Na Li, Yong Zhao.  (2025)  A fluorescent fiber pH sensor for sensitive, wide-range detection using orange-emitting silicon quantum dots.  ANALYTICA CHIMICA ACTA,      [PMID:40670006] [10.1016/j.aca.2025.344330]

99. Xiongchao Liu, Xinyu Li, Na Gao, Zhiwei Liu, Yuangang Li, Mingli Wang, Tifeng Jiao.  (2025)  Langmuir-Blodgett composite films of amphiphilic perylene diimide and naphthalene diimide derivatives for acid-base gas sensing and surface-enhanced Raman scattering.  Sustainable Materials and Technologies,      [PMID:] [10.1016/j.susmat.2025.e01707]

100. Jun Zeng, Tao Jia, Tong Chen, Huilin Liu, Zhaojie Sun, Xiaoyong Huang, Yunfei Shang, Shuwei Hao, Chunhui Yang.  (2025)  Excitation-modulated upconversion nanoprobes for switchable imaging and phototherapy.  Applied Materials Today,      [PMID:] [10.1016/j.apmt.2025.102979]

101. Xiaoman Huang, Zixuan E, Hao Yang, Guihong Lai, Shuang Liu, Shujia Liao, Jiaqi Lin, Siyi Liu, Chen Long, Haoran Lin, Danqi Lu, Shuisheng Li, Yong Zhang.  (2025)  Immunological Characteristics of Triploid Grass Carp (Ctenopharyngodon idella) and evaluation of its phagocytosis in the macrophages of head kidney.  FISH & SHELLFISH IMMUNOLOGY,      [PMID:41205782] [10.1016/j.fsi.2025.110987]

102. Jie Song, Qian Pu, Chen Chen, Xingcheng Liu, Xinlei Zhang, Zejun Wang, Jin Yan, Xuedong Wang, Huili Wang, Qiuhui Qian.  (2025)  Neurological Outcomes of Joint Exposure to Polystyrene Micro/Nanospheres and Silver Nanoparticles in Zebrafish.  ENVIRONMENTAL HEALTH PERSPECTIVES,  133  (5): (057007).  [PMID:40138633] [10.1289/EHP14873]

103. Zhaokang Zheng, Zhen Liu, Yujie Ma, Aiwu Wang, Cangtao Zhou, Muhammad Humayun, Mohamed Bououdina.  (2025)  Oxygen-vacancy-rich MoO3-x nanostructures for ultrahigh cationic dye adsorption: Kinetics, isotherms, thermodynamics, and mechanism.  DYES AND PIGMENTS,      [PMID:] [10.1016/j.dyepig.2025.113454]

104. Yongfei Zhu, Jiayi Li, Shujuan Yu.  (2025)  Robust superhydrophilic/underwater superoleophobic cotton fabric decorated by β-FeOOH and PVA with excellent oil-water separation and photodegradation properties.  SEPARATION SCIENCE AND TECHNOLOGY,      [PMID:] [10.1080/01496395.2025.2595175]

105. Hui Ying Bai, Rui Hao Chen, Iek Man Lei, Zhi Jian Wang, Zi Liang Wu.  (2026)  Marangoni-Effect-Driven Locomotion of Patterned Gels with Evolved Shape Changes and Varied Motion Modes.  ACS Applied Materials & Interfaces,      [PMID:41555737] [10.1021/acsami.5c24354]

106. Qin Fang, Mengdie Qin, Ke-jun Tan.  (2026)  One-step synthesis of multicolor fluorescent carbon dots from neutral red for applications in LEDs and fingerprint detection.  DYES AND PIGMENTS,      [PMID:] [10.1016/j.dyepig.2026.113695]