储存缓冲液：无水DMSO
Ready-to-use SYTOGreen 9 Live Cell Nucleic Acid Stain是一款即用型、膜通透的绿色荧光核酸染料，无需DMSO溶解或浓度校准，专为原核与真核生物的核酸可视化设计。核心原理染料分子可穿透完整细胞膜进入活细胞内，与DNA/RNA特异性嵌入结合后荧光信号显著增强 (未结合时量子产率＜0.01，结合后提升至0.4以上)，通过荧光强度即可直观反映样本中核酸含量及生物活性。其光学特性优异，最大激发波长为485 nm，最大发射波长为 498 nm，适配常规荧光检测设备的FITC通道，荧光强度与核酸含量线性相关。可兼容流式、荧光显微镜及酶标仪等多种平台，实现微生物死活区分、活细胞实时成像、增殖监测等一站式核酸可视化应用。
在生命科学研究与工业检测中，“快速、精准、便捷地获取核酸及生物活性信息”是核心需求。Ready-to-use SYTOGreen 9的推出有效解决，其“即用型”特性大幅缩短实验准备时间，以低毒强荧光活性的优势实现了“活细胞保护”与“高灵敏检测”的兼顾，而与多种实验技术的兼容性则打破了不同场景下染料更换的壁垒。在微生物学领域，其独特的膜渗透特性使其可与碘化丙啶 (PI)联合应用快速区分食品、水体中的活菌与死菌鉴别，替代耗时的培养法；在细胞生物学中，为活细胞动态监测提供了稳定的荧光标记工具，是推动核酸检测向“高效化、便捷化、通用化”升级的关键试剂。 仅限体外科研用途 不得用于临床诊断或治疗。
注：效能等同Thermo Fisher SYTO® 9-Fluorescent Nucleic Acid Stains (S34854)。
SYTO® 为 Thermo Fisher 的商标及注册商标。
应用范围：
1、微生物学应用：革兰氏阳性菌/阴性菌死活区分、真菌活性检测、水体/食品微生物污染快速筛查、生物膜中活菌定量；
2、细胞生物学应用：哺乳动物活细胞荧光成像、细胞增殖与凋亡动态监测、干细胞分化过程中核酸含量追踪、药物对细胞代谢的影响评估；
3、工业质控应用：发酵液中微生物活性监测、化妆品/药品微生物限度快速检测、环境样本中病原微生物筛查；
产品特点：
1、荧光信号增强显著：结合核酸后荧光增强超40倍，摩尔吸光系数>60,000 cm⁻¹M⁻¹；
2、低细胞毒性：毒低浓度下不影响细胞活性，适合活体样本长期动态观察；
3、适用范围广泛：兼容流式细胞仪、荧光显微镜等多种检测设备，无需专属仪器；
4、联合兼容性好：可与PI等膜非透性染料高效搭配，实现活/死细胞 (细菌)精准区分，无信号交叉干扰；
5、操作简便高效：无需固定、破膜或洗涤步骤，直接孵育15-20 min即可上机，大幅缩短实验周期；
产品参数：
1、结合 DNA Ex/Em：485/498 nm
2、结合 RNA Ex/Em：486/501 nm
光谱特性 (SYTO9与DNA结合的吸收/发射光谱)：
产品组分：
| 组分 | 规格 (100 T) | Ready-to-use SYTOGreen 9 Live Cell Nucleic Acid Stain (5 mM)
| 100 μL |
|
注：使用次数按照细菌染色，染色液体积1 mL浓度5 μM计算。
注意事项：
1、避光防反复冻融：首次开封按单次用量分装，−20 ℃避光保存。
2、少液先离心：余量较少时，建议使用前先快速离心数秒钟，使液体充分沉降再取。
3、塑料容器稀释：染料易吸附玻璃材质的容器，全程用 PP/PE 管。
4、远离淬灭剂：禁用含叠氮化物、＞10 mM EDTA 或巯基缓冲液；优选无磷酸盐缓冲液。
5、去污剂限量：SDS/Triton X-100 终浓度≤0.1 %，避免＞0.5 % 破坏染料结构。
6、器皿洁净：温和洗涤剂→热水→去离子水各冲洗数次，杜绝残留荧光背景。
7、操作避强光：配液、染色、保存全程避自然光及强LED，确保信号稳定。
8、本产品仅限于科研用途并且不得存放于普通住宅内。
9、为了您的安全和健康，请遵循您所在常规实验室安全规定。
使用说明：
1、SYTOGreen 9进行真核细胞染色时，除细胞核会被染色之外，细胞质通常也会呈现弥散性标记信号，核小体会呈现出较高的荧光标记效果。
2、SYTOGreen 9具有穿透细胞膜的能力，在中性pH条件下带净正电荷，可能会对线粒体染色。染色活酵母菌的结果就是主要染色菌体内的线粒体。
3、本产品为浓度为5 mM的DMSO溶液，使用时用适宜的缓冲液稀释至工作浓度孵育染色即可。
4、SYTOGreen 9染色浓度和时间建议根据不同的样本类型进行调整，推荐染色条件：
| 样品类型 | SYTOGreen 9染色浓度 | 染色条件 | | 细菌 | 50 nM-20 μM | 孵育时间 1 min-30 min | | 真核细胞 | 10 nM-5 μM | 孵育时间 10 min-120 min | | 微阵列 | 50 nM（TE 缓冲液） | 孵育时间 5 min，冲洗后晾干 |
|
常见疑问与解答：
1、问：SYTOGreen 9与常用的DAPI和Hoechst33342有什么区别？
答：SYTOGreen 9膜透性更强、适配样本更广 (尤其细菌)、兼容多检测平台，而DAPI和Hoechst33342侧重DNA特异性结合、低背景，更适合真核细胞细胞核染色或固定样本分析：
| 对比 | SYTOGreen 9
| DAPI | Hoechst33342 | | 染色特异性 | 结合DNA和RNA无序列 (偏好) | 特异性结合DNA的AT富集区 (不结合RNA) | 特异性结合DNA的AT富集区 (不结合RNA) | | 膜透性 | 穿透活菌/死菌、细菌/真核细胞，无需破膜 | 活体真核细胞穿透性差，多需固定+破膜；细菌需透化处理 | 可穿透活体真核细胞，细菌穿透性弱 | | 激发光与荧光强度 | 蓝光激发，结合核酸后荧光增强约40倍 | 紫外激发，结合核酸后荧光增强约20倍 | 紫外激发，结合核酸后荧光增强约10倍 | | 细胞毒性 | 低浓度下不影响细胞活性，适合活体长时间观察 | 紫外激发+染料本身有一定毒性，不适合活体长期培养 | 毒性低于DAPI，可用于活体染色但需控制浓度 | | 适用样本类型 | 细菌、真菌、酵母、动植物细胞 (活菌/固定) | 固定细胞、组织切片、细胞核染色 (细菌需透化处理) | 活体/固定真核细胞、细胞核染色 (细菌应用少) |
|
2、问：为什么SYTOGreen 9对不同细菌染色效果差异很大？
答：不同细菌染色效果不同受到多种因素的影响，主要因素有以下几个：革兰氏阴性菌会由于菌体的外膜屏障染色效果通常弱于革兰氏阳性菌；细菌生长的状态和所处的生长阶段影响也比较大，对数期的细菌染色效果最好；不同菌种的最适染料浓度不一样。
Ready-to-use SYTOGreen 9 Nucleic Acid Stain is a ready-to-use, membrane-permeable green fluorescent nucleic acid dye that requires no DMSO solubilization or concentration calibration, specially designed for nucleic acid visualization in prokaryotes and eukaryotes. Core Principle: The dye molecules can penetrate intact cell membranes into viable cells and intercalate specifically with DNA/RNA, resulting in a significant enhancement of fluorescent signal (quantum yield ＜0.01 in the unbound state, increased to over 0.4 in the bound state). The nucleic acid content and biological activity in samples can be directly reflected by the fluorescence intensity. It features excellent optical properties with an excitation maximum at 485 nm and an emission maximum at 498 nm, matching the FITC channel of conventional fluorescence detection equipment, and its fluorescence intensity has a linear correlation with nucleic acid content. Compatible with flow cytometry, fluorescence microscopy, microplate readers and other platforms, it enables one-stop nucleic acid visualization applications such as microbial viability discrimination, real-time live cell imaging and proliferation monitoring.
In life science research and industrial testing, the core demand is to obtain nucleic acid and biological activity information rapidly, accurately and conveniently, which is effectively addressed by the launch of Ready-to-use SYTOGreen 9. Its ready-to-use property greatly shortens experimental preparation time; with the advantages of low toxicity and strong fluorescent activity, it achieves both viable cell protection and high-sensitivity detection; its compatibility with a variety of experimental technologies breaks the barrier of dye replacement in different scenarios. In the field of microbiology, its unique membrane-permeable property allows it to be used in combination with Propidium Iodide (PI) for rapid discrimination of viable and dead bacteria in food and water samples, replacing time-consuming culture methods. In cell biology, it provides a stable fluorescent labeling tool for dynamic monitoring of live cells, serving as a key reagent to upgrade nucleic acid detection towards high efficiency, convenience and universal applicability. For in vitro research use only. Not for clinical diagnosis or treatment.
Note: Performance equivalent to Thermo Fisher SYTO® 9-Fluorescent Nucleic Acid Stain (S34854).SYTO® is a trademark and registered trademark of Thermo Fisher.
Application Scope
1. Microbiology Applications: Viability discrimination of Gram-positive/Gram-negative bacteria, fungal activity detection, rapid screening of microbial contamination in water/food, quantification of viable bacteria in biofilms;
2. Cell Biology Applications: Fluorescent imaging of mammalian live cells, dynamic monitoring of cell proliferation and apoptosis, tracking of nucleic acid content during stem cell differentiation, evaluation of drug effects on cell metabolism;
3. Industrial Quality Control Applications: Monitoring of microbial activity in fermentation broth, rapid detection of microbial limits in cosmetics/pharmaceuticals, screening of pathogenic microorganisms in environmental samples.
Product Features
1. Significant fluorescent signal enhancement: Over 40-fold fluorescence enhancement upon nucleic acid binding, molar extinction coefficient >60,000 cm⁻¹M⁻¹;
2. Low cytotoxicity: No impact on cell activity at low concentrations, suitable for long-term dynamic observation of living samples;
3. Wide applicability: Compatible with various detection equipment including flow cytometers and fluorescence microscopes, no exclusive instrument required;
4. Excellent combinatorial compatibility: Can be efficiently combined with membrane-impermeable dyes such as PI to achieve accurate discrimination of live/dead cells (bacteria) without signal crosstalk;
5. Simple and efficient operation: No fixation, membrane lysis or washing steps required; direct incubation for 15-20 min before loading, greatly shortening the experimental cycle.
Product Parameters
1. Ex/Em upon DNA binding: 485/498 nm
2. Ex/Em upon RNA binding: 486/501 nm
Spectral Characteristics (Absorption/Emission Spectrum of SYTO9-DNA Complex)
Product Components
| Component | Specification (100 Tests) | | Ready-to-use SYTOGreen 9 Nucleic Acid Stain (5 mM) | 100 μL |
|
Note: The number of uses is calculated based on bacterial staining with 1 mL staining solution at a concentration of 5 μM.
Precautions
1. Protect from light and avoid repeated freeze-thaw: Aliquot into single-use volumes upon first opening and store at −20 ℃ in the dark.
2. Centrifuge before use if volume is low: For small remaining volumes, brief centrifugation for a few seconds before use is recommended to ensure complete sedimentation of the solution prior to pipetting.
3. Dilute in plastic containers: The dye is prone to adsorption to glass containers; use PP/PE tubes throughout the process.
4. Avoid quenchers: Do not use buffers containing azides, EDTA ＞10 mM or thiols; phosphate-free buffers are preferred.
5. Limit detergent concentration: Final concentration of SDS/Triton X-100 ≤0.1 %; avoid concentrations ＞0.5 % which may damage the dye structure.
6. Clean vessels thoroughly: Rinse several times with mild detergent → hot water → deionized water to eliminate residual fluorescent background.
7. Avoid strong light during operation: Protect from natural light and strong LED light during solution preparation, staining and storage to ensure stable signal.
8. This product is for research use only and shall not be stored in ordinary residential premises.
9. For your safety and health, comply with the general laboratory safety regulations of your institution.
Instructions for Use
1. When staining eukaryotic cells with SYTOGreen 9, in addition to nuclear staining, the cytoplasm usually shows a diffuse labeling signal, and nucleoli exhibit a strong fluorescent labeling effect.
2. SYTOGreen 9 is membrane-permeable and carries a net positive charge under neutral pH conditions, which may result in mitochondrial staining. For example, staining of viable yeast cells primarily labels mitochondria within the cells.
3. This product is a 5 mM DMSO solution; dilute to the working concentration with an appropriate buffer and incubate for staining before use.
4. The staining concentration and incubation time of SYTOGreen 9 are recommended to be adjusted according to different sample types. Recommended staining conditions:
| Sample Type | SYTOGreen 9 Staining Concentration | Staining Conditions | | Bacteria | 50 nM-20 μM | Incubation: 1 min-30 min | | Eukaryotic Cells | 10 nM-5 μM | Incubation: 10 min-120 min | | Microarray | 50 nM (in TE buffer) | Incubation: 5 min, rinse and air-dry |
|
Frequently Asked Questions (FAQs)
1. Q: What is the difference between SYTOGreen 9 and the commonly used DAPI and Hoechst33342?
A: SYTOGreen 9 has stronger membrane permeability and wider sample applicability (especially for bacteria), and is compatible with multiple detection platforms. In contrast, DAPI and Hoechst33342 focus on specific DNA binding with low background, making them more suitable for nuclear staining of eukaryotic cells or analysis of fixed samples.
| Comparison Index | SYTOGreen 9
| DAPI | Hoechst33342 | | Staining Specificity | Binds DNA and RNA without sequence preference | Specifically binds to AT-rich regions of DNA (no RNA binding) | Specifically binds to AT-rich regions of DNA (no RNA binding) | | Membrane Permeability | Penetrates viable/dead bacteria and eukaryotic cells, no membrane lysis required | Poor permeability to live eukaryotic cells (fixation + membrane lysis mostly required); bacterial permeabilization needed | Penetrates live eukaryotic cells, weak permeability to bacteria | | Excitation Light & Fluorescence Intensity | Blue light excitation, ~40-fold fluorescence enhancement upon nucleic acid binding | UV light excitation, ~20-fold fluorescence enhancement upon nucleic acid binding | UV light excitation, ~10-fold fluorescence enhancement upon nucleic acid binding | | Cytotoxicity | No impact on cell activity at low concentrations, suitable for long-term observation of living samples | UV excitation + inherent dye toxicity, not suitable for long-term culture of living samples | Lower toxicity than DAPI, applicable for live cell staining but concentration needs to be controlled | | Applicable Sample Types | Bacteria, fungi, yeast, animal and plant cells (viable/fixed) | Fixed cells, tissue sections, nuclear staining (bacterial permeabilization needed) | Viable/fixed eukaryotic cells, nuclear staining (rarely used for bacteria) |
|
2. Q: Why is the staining effect of SYTOGreen 9 significantly different for various bacteria?
A: The differential staining effect on various bacteria is affected by multiple factors, the main ones being as follows: Gram-negative bacteria usually show weaker staining than Gram-positive bacteria due to the outer membrane barrier of bacterial cells; the growth state and phase of bacteria also have a significant impact, with the best staining effect observed in logarithmic-phase bacteria; the optimal dye concentration varies among different bacterial strains.
ZJ26F0939390
危险品化学品经营许可证（带存储）