仪器标准品-Instrument Standards

我们为细胞分析仪、粒度仪、流式细胞仪和荧光显微镜等分析仪器制造全面的标准品。我们的目录包括 NIST Traceable Particle Size Standards、SureCount™ Count Standards、ViaCheck™ Cell Viability and Concentration Controls，以及用于流式细胞术的各种荧光和抗体捕获珠以及用于成像的荧光珠和载玻片。

DESCRIPTION

Right Reference Standard™ microspheres are suspensions of highly uniform microspheres labeled with a single fluorochrome at a high intensity level. These
microspheres exhibit excitation and emission spectra matching those of cell samples labeled with the same fluorochromes and approximate the size of
human lymphocytes (7-9µm). Right Reference Standard™ microspheres are suspended in a sterile-filtered, isotonic, buffered solution (pH 7.4). Quantum™
QC or Quantum™ MESF microspheres may be used for applications requiring multiple intensities.

Right Reference Standard™ 微球是高度均匀的微球悬浮液，以高强度水平标记有单一荧光染料。这些微球的激发和发射光谱与用相同荧光染料标记的细胞样品相匹配，大小接近
人淋巴细胞 (7-9µm)。 Right Reference Standard™ 微球悬浮在无菌过滤的等渗缓冲溶液 (pH 7.4) 中。量子™QC 或 Quantum™ MESF 微球可用于需要多种强度的应用。

CHARACTERISTICS

Mean Diameter: 7-9µm
Particle Concentration: 2 x 106
microspheres/mL
MATERIAL
Material Supplied
Right Reference Standard™ microspheres
Material Required
Cell suspension solution
Appropriate sized test tubes
Flow cytometer

PROCEDURE

Researchers are advised to optimize the use of particles in any application.
Determining Instrument Specific Target Channels
1. On the cytometer, acquire a stained sample that is representative (e.g., a positive control) of the samples you wish to analyze. Adjust settings as needed
to have the sample fall into the desired range.
2. Prepare Right Reference Standard microspheres by lightly shaking bottle (do not sonicate or vortex).
3. Add a drop (~ 50 ul) to 400 ul of buffer for analysis. Using the same settings as for the positive sample in step 1, gate on the singlet population
of microspheres in a bivariant histogram (dot plot) showing side scatter (SSC) vs. forward scatter (FSC) (as shown above) and apply to a
fluorescence histogram.
4. Repeat runs 3 times with independent prepared samples for determination of inherent run variability.
5. Use the mean value of these runs as your instrument specific target channel. Outliers should be discarded and the run repeated.
Note: Initial settings may need to be adjusted to fit both the positive control and reference standard within the same window of analysis. If adjustments are
made, record the new instrument specific target channel.
Subsequent Daily Set-up and Quality Control
1. Set the PMT and compensations to values used in the previous section.
2. Run the Right Reference Standard to ensure that they fall near the instrument specific target channels. Tolerance on absolute differences between the
set target channel value and the measured channel value will need to be determined by the user, we encourage the use of Levey-Jennings charts to
assist with this, as well as the use of daily QC/setup beads.
Expected Results
The Right Reference Standard™ microspheres are labeled with a single fluorochrome. The microspheres will be visible as an intense fluorescent peak in the
applicable fluorescent channel.

建议研究人员在任何应用中优化粒子的使用。
确定仪器特定的目标通道
1. 在流式细胞仪上，采集代表您要分析的样本（例如阳性对照）的染色样本。根据需要调整设置使样本落入所需范围。
2. 轻轻摇晃瓶子（不要超声或涡旋）制备正确的参考标准微球。
3. 向 400 ul 缓冲液中加入一滴（~ 50 ul）进行分析。使用与步骤 1 中正样本相同的设置，对单线群体进行门控显示侧向散射 (SSC) 与前向散射 (FSC)（如上所示）的双变量直方图（点图）中微球的数量，并适用于荧光直方图。
4. 用独立制备的样品重复运行 3 次，以确定固有的运行变异性。
5. 使用这些运行的平均值作为仪器特定的目标通道。应丢弃异常值并重复运行。
注意：可能需要调整初始设置以适应同一分析窗口内的阳性对照和参考标准。如果调整是
制作，记录新仪器特定的目标通道。随后的日常设置和质量控制
1. 将 PMT 和补偿设置为上一节中使用的值。
2. 运行正确的参考标准，确保它们落在仪器特定目标通道附近。公差之间的绝对差异设置目标通道值和测量通道值需要用户确定，我们鼓励使用 Levey-Jennings 图协助解决此问题，以及使用日常 QC/设置珠。
预期成绩
Right Reference Standard™ 微球标有单一荧光染料。微球将作为强烈的荧光峰出现在适用荧光通道。

NOTES

Prior to acquiring the Right Reference Standard™ microspheres, the flow cell should be free of debris. This can be accomplished by running a 10% solution
of household bleach (follow instrument manufacturer’s recommendations) for 5 minutes, followed by distilled water for another 5 minutes at the highest flow
rates allowable (will assist in clearance). Should this fail, follow the following steps:
• Run the cytometer’s debubble/purge cycle.
• Verify that the instrument is properly compensated.
• Perform Manufacturer’s recommended monthly cleaning procedure.
• Check the properties of diluent and sheath fluid (e.g., especially changes in pH).
• Check alignment of the instrument.
• Prepare a new sample and run once again.

在获取 Right Reference Standard™ 微球之前，流通池应该没有碎屑。这可以通过运行 10% 的解决方案来完成家用漂白剂（按照仪器制造商的建议）5 分钟，然后以最高流量再用蒸馏水 5 分钟
允许的利率（将有助于清关）。如果此操作失败，请执行以下步骤：
• 运行细胞仪的除泡/清除循环。
• 验证仪器是否得到适当补偿。
• 执行制造商推荐的每月清洁程序。
• 检查稀释剂和鞘液的性质（例如，特别是 pH 值的变化）。
• 检查仪器的对齐情况。
• 准备一个新样品并再次运行。

REFERENCES

1. Schwartz, A., E. Fernandez-Repollet. 1993. Development of clinical standards for flow cytometry. Clinical Flow Cytometry, Ann NY Acad Sci. 677: 28-39.
2. Shapiro, H.M. 1995. Practical flow cytometry, 3rd ed. New York: Wiley Liss, Inc.
TRADEMARKS AND REGISTERED TRADEMARKS
Right Reference Standard™ is a trademark of Bangs Laboratories, Inc.
Cy™, including Cy5, is a trademark of GE Healthcare Limited. These products are manufactured under license from Carnegie Mellon University under U.S.
Patent Number 5,268,486 and related patents.
STORAGE AND STABILITY
Store at 2-8°C. Freezing may result in irreversible aggregation. Stable for 6 or 12 months from date of purchase, as noted on the product label, provided the
product is handled in accordance with the manufacturer’s recommendations. The reagent should be kept in is opaque bottle.
This product is for research use only and is not intended for use in humans or for in vitro diagnostic use.

1. Schwartz, A., E. Fernandez-Repollet。 1993. 流式细胞仪临床标准的发展。临床流式细胞术，Ann NY Acad Sci。 677：28-39。
2.夏皮罗，H.M. 1995. 实用流式细胞术，第 3 版。纽约：威利利斯公司
商标和注册商标
Right Reference Standard™ 是 Bangs Laboratories, Inc. 的商标。
Cy™，包括 Cy5，是 GE Healthcare Limited 的商标。这些产品是在美国卡内基梅隆大学的许可下生产的。
专利号 5,268,486 及相关专利。
储存和稳定性
储存在 2-8°C。冷冻可能导致不可逆的聚集。如产品标签上所述，自购买之日起可稳定保存 6 或 12 个月，前提是
产品按照制造商的建议进行处理。试剂应保存在不透明的瓶子中。
本产品仅供研究使用，不可用于人体或体外诊断用途。

ORDERING INFORMATION

Cat. Code Description Sizes
512 Right Reference Standard™ Fluorescein, High Intensity 5mL
515 Right Reference Standard™ Phycoerythrin, High Intensity 5mL
518 Right Reference Standard™ PE-Cy™5, High Intensity 5mL
521 Right Reference Standard™ APC, High Intensity 5mL