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View additional product information for Attune™ NxT Flow Cytometer, blue/violet/yellow - FAQs (A24859)
51 product FAQs found
不一定。许多用户使用未染色细胞结合FMO对照，来确定他们的阳性群体。
有许多因素会引起样品率低下：
•系统可能堵塞
•阙值水平可能设置的太高
•阙值参数的PMT电压可能设置的太低
•样品可能被过分稀释或是没有充分混匀
•样品注射器可能松动
有许多因素会引起样品率高：
•可能流动细胞中有气泡
•阙值水平可能设置的太低
•阙值参数的PMT电压可能设置的太高
•样品浓度可能太高
•样品流速可能过高
•样品可能被细菌污染
打开Attune NxT软件，打开主菜单，进入“仪器”选项。最左边的选项是“系统日志”。点击该选项，即可打开日志文件。页面的底部有一个标有“导出”的按钮。使用该按钮可导出并保存日志文件。
我们建议查看箱子内部，可能在缓冲液袋下方。
请发送邮件至细胞分析技术支持部门(technical-molecularprobes@lifetech.com)，请在邮件的主题行输入“flow”或“Attune”以确保邮件能够顺利投递。请在邮件中描述您的应用类型，实验方法以及结果，最好能提供得到数据的样品供我们分析。
如果明显是仪器问题，请附上您的仪器序列号发邮件至仪器服务部门instrumentservices@thermofisher.com。如果是软件或者应用相关问题，或者您不清楚问题的原因，请发送邮件至细胞分析技术支持部门technical-molecularprobes@lifetech.com——请在邮件的主题行输入“flow”或“Attune” 以确保邮件能够顺利投递。
关于流式细胞术抗体组合的设计，见我们新的分子探针流式细胞术抗体组合设计工具。抗体组合设计工具可以帮助您经过简单的几步，为您的流式细胞染色组合选择荧光抗体偶联物：选择抗体种属反应性，最多选择14个目标靶标（包括活性染料），选择激光或者您希望观察的荧光。
这取决于您的仪器。人们已经试验了17种颜色（Perfetto SP, Chattopadhyay PK, and Roederer M (2004) Seventeen-colour flow cytometry: unraveling the immune system. Nat Rev Immunol 4:648–655.）或更多，但是这需要很多规划和测试使得其能够起作用。Attune NxT声波聚焦流式细胞仪可以运行14种颜色，前向和侧向散射，其配备了4种激光（紫、蓝、黄和红）。
FMO对照（染色减一）是一种采用扣除一个颜色的所有染色组合在来标记细胞或微珠的对照。请针对您的染色组合准备一份FMO对照。这些对照对合理设置您的数据门限很有帮助。
这些同型对照并不是非特异性结合的最适宜的对照，同型对照的使用正渐渐的离开流式细胞领域。作为替代，可使用未染色细胞确定阴性群体，使用单独染色对照设置补偿，使用减一染色（FMO）对照设置区域和门限。
您可能需要考虑是否有必要使用同型对照。以下是您可能感兴趣的部分参考文献：
•O'Gorman MR, Thomas J. (1999) Isotype controls—time to let go? Cytometry 38:78–80.
•Keeney M, Gratama JW, Chin-Yee IH et al. (1998) Isotype controls in the analysis of lymphocytes and CD34+ stem/progenitor cells by flow cytometry—time to let go! Cytometry 34:280–283.
•Hulspas R, O'Gorman MR, Wood BL et al. (2009) Considerations for the control of background fluorescence in clinical flow cytometry. Cytometry B Clin Cytom 76:355–364.
•Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline—Second Edition. Clinical and Laboratory Standards Institute, (CLSI). Document H42-A2 Volume 27 No.16, 2007.
•Clinical Flow Cytometric Analysis of Neoplastic Hematolymphoid Cells; Approved Guideline—Second Edition. Clinical and Laboratory Standards Institute, (CLSI). Document H43-A2 Volume 27 No. 11, 2007.
我们的流式细胞仪大小校准试剂盒含有无荧光微粒大小的校准标准品，能够简便、精确地通过流式细胞仪测定细胞大小。此试剂盒包含了六种悬浮的高度均一的聚苯乙烯微球，直径分别为1.0 µm、2.0 µm、4.0 µm、6.0 µm、10.0 µm和15.0 µm。所有微球悬液以方便的滴管形式提供。此试剂盒通过大小已知的非荧光微球的散射特性，大致测定细胞大小。
Attune NxT声波聚焦流式细胞仪检测的下限是0.5 µm。
Attune NxT自动进样器是Attune NxT声波聚焦流式细胞仪的可选附件，能够快速分析至多384份样品。对于不同形式平板——无论96孔还是384孔板——它都具有广泛的兼容性。内置智能探针，最大限度避免堵塞和截留（<0.5%），防止损坏仪器。该仪器通过通气而不是振荡混合，确保样品的均质性，同时维持细胞的活性。它还可作为Attune NxT流式细胞仪关机程序的一个步骤来执行自动清洗。无论采用何种取样方法（试管或平板）和数据采集速率，它都能提供一致的数据。
•模块化设计—多种配置可选，可现场升级。
•节省时间—在不降低数据质量的前提下，速度提高10倍以上。
•简化样品准备流程—无需洗涤，无裂解选项，流体不堵塞。
•可实现独特的应用—可针对广泛的细胞类型和样本构建复杂的实验方案。
针对多参数分析应用，Attune NxT声波聚焦流式细胞仪可以选择最多4个激光器和14色配置，采用模块化系统设计来满足大部分试验需求和实验室预算。创新的光路设计，有助于确保四个空间分隔的激光器精确对齐样品流，确保长期的数据一致性、优越的性能和出色的可靠性。仪器可配备多达4个具有平顶光束的固态激光器（405 nm、488 nm、561 nm和637 nm）。
Attune NxT声波聚焦流式细胞仪的声波聚焦技术采用超声波辐射压力（>2 MHz），将微粒运送至样品流中心。之后，这股预聚焦流注入到为样品提供额外流体动力学压力的鞘液流中。这种“声波辅助流体动力学聚焦”技术可以在任何进样速率的情况下都保证样品流中心范围集中、激光照度均一。在传统的依赖于流体动力学聚焦技术的流式细胞仪中，样品中心需要变宽来适应流速的增加，从而导致激光照度不均一。
细胞计数是一种测量细胞和微粒物理化学特性的方法。流式细胞术可在流式细胞仪内使细胞或微粒单独通过激光来测定其特性，它可用于测定单细胞，针对样品中的每个细胞提供独立的测定结果。此外，还可提供这些样品特性的统计分布结果。 流式细胞仪由三个子系统组成：流体系统、光学系统和电子系统。流体系统负责使细胞移动，从而对其进行分析。光学系统负责产生和收集光信号。电子系统负责将合适的光学信号转化为电子信号，以便进行计算机分析。
To minimize the number of failed performance tests on your Attune NxT Flow Cytometer, we suggest the following:
• Make sure to carry out daily, weekly, and monthly maintenance procedures (pages 8-14 of the Maintenance and Troubleshooting Guide, linked below).
• Always run 3 startup cycles and 2 Rinse procedures (pages 46-47 of the Maintenance and Troubleshooting Guide linked below).
• If the performance test fails, carry out the following priming sequence: run 3 startup cycles, 2 de-bubble cycles, and 5 rinse procedures, followed by running the SIP sanitize function.
Please review the Attune Flow Cytometry Maintenance and Troubleshooting Guide for more information about performing routine maintenance procedures and troubleshooting.
Find additional tips, troubleshooting help, and resources within our Flow Cytometry Support Center.
Yes. The Attune Nxt Flow Cytometer can detect down to 0.2 µm and using a special configuration, down to 0.1 µm. Optimize your settings using Flow Cytometry Sub-micron Particle Size Reference Kit, Cat. No. F13839.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Yes. You would need to purchase Attune NxT Custom Filter Holder Kit, Cat. No. A27784 to make your own custom emission or dichroic filters in order to customize the optical configuration of the instrument. Each kit contains sufficient parts to make 2 emission filters and 2 dichroic filters. The kit includes instructions for attaching the filter to the filter blade, but does not include the glass filter. Other accessories to optimize for special applications include Cat No. 100083194 Attune NxT Small Particle Side-Scatter Filter, Cat. No. 100083194 and Attune NxT Blocker Bar Conversion Kit, Cat No. A35966.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Yes. There are 6 lasers to choose from (blue, green, yellow, red, violet, and violet 6). You can start with one laser and at any time upgrade the same instrument with up to 4 lasers to utilize a maximum of 14 detection channels. Using the Violet 6 laser along with the blue, red, and yellow lasers, one can detect up to 16 channels.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Because of the acoustic-assisted hydrodynamic technology, the Attune NxT Flow Cytometer uses only 1/10th the amount of fluids, and is 10X faster and less sensitive to clogging compared to standard flow cytometers. You can set the flow rate from 12.5-1,000 µL/min and collect up to 65,000 events/sec using from 20 µL to 4 mL of sample volume. These unique properties allow for rare event analysis, as well standard flow analysis.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Filters are recommended for replacement once every three months, unless performing small particle detection when once-a-month replacement is recommended.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
In an emergency, we recommend using the highest purity of water available in place of the shutdown solution and then switching to the Attune Shutdown Solution as soon as possible.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
In an emergency, we recommend using the highest purity of water available in place of the wash solution and then switching to the Attune Wash Solution as soon as possible.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
You can tell whether the compensation data are included by looking for the $SPILL or $COMP keyword embedded within the FCS files. You can also see the FCS file information within the Attune NxT software by going to the "View" tab and checking the box labeled "FCS File Panel". This will open a window which shows the information contained in the FCS file associated with the selected sample. It will include any compensation ($SPILLOVER) that has been done.
Step 2 allows for antibodies you have already purchased or are from a vendor outside Thermo Fisher Scientific. If you are undecided about which format you want for something, you can add the antigen name in the "Antibodies you need" section and then choose a placeholder fluorochrome in the following step. Both will help you keep track of channels that are reserved for reagents that you do not plan to buy from Thermo Fisher Scientific.
There are both PDF (printable) and CSV (spreadsheet) exports available in the summary step for you to take away for reference and ordering purposes.
Yes, you can define a new cytometer using the "Enter your cytometer manually" link below the cytometer dropdown menu.
Select a cytometer from the dropdown menu and then select the "Edit cytometer settings" link to modify the configuration as needed.
Each dot indicates a different fluorochrome that Thermo Fisher Scientific has available in each channel of the cytometer you have chosen.
You may export the spreadsheet found on step 5 and share it.
Select your target species first, and then type your antigen. A drop-down list will appear. Select the antigen from the list and click on "Confirm selections." If you have an antibody that is not on the list, in the "Antibodies you already have" section, you can enter a name and select "use this name anyway."
Yes, the previous work is saved. If you plan to leave the Panel Builder and come back later, be sure to select "Save" in the lower right corner to save your panel.
No. Many users are using unstained cells in combination with FMO controls to identify their positive populations.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
There are several things that can cause a low event rate:
-The system may be clogged
-The threshold level may be set too high
-The PMT voltage for the threshold parameter may be set too low
-The sample may be too dilute or not adequately mixed
-The sample syringe may be loose
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
There are several things that can cause a high event rate:
-There may be an air bubble in the flow cell
-The threshold level may be set too low
-The PMT voltage for the threshold parameter may be set too high
-The sample may be too concentrated
-The sample flow rate may be too high
-The sample may have a bacterial contamination
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Open the Attune NxT software, open the main menu, and go to the “Instrument” tab. The option on the far left is “System Log”. Click on that and it will open the log file. At the bottom of the page there is a button labeled “Export”. Use this to export and save the log file.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
We recommend looking inside the box, possibly underneath the bag of buffer.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Send an email to Cellular Analysis Technical Support at technical-molecularprobes@lifetech.com, including the words “flow*#148; or “Attune” in the subject line for proper routing of your email. Please describe the type of application you are doing, how you set up the experiment, and your results. It would be ideal if you could send us a sample of your data that we can review.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
If the issue is clearly an instrument problem, send an email to Instrument Services at instrumentservices@thermofisher.com with your instrument serial number. If the issue is software or application-related, or if you do not know the source of the problem, send an email to Cellular Analysis Technical Support at technical-molecularprobes@lifetech.com, including the words “flow*#148; or “Attune” in the subject line for proper routing of your email.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
For help with designing panels for flow cytometry, see our new Molecular Probes Flow Cytometry Panel Design Tool (http://www.thermofisher.com/us/en/home/life-science/cell-analysis/flow-cytometry/flow-cytometry-panel-design-tool.html) The panel design tool can help you choose fluorescent antibody conjugates for your flow cytometry panel in a few easy steps: pick the antibody species reactivity, select up to 14 targets of interest (choices include viability dyes), and choose the lasers or fluorophores you want to view.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
It depends on your instrument. People have done 17 colors (Perfetto SP, Chattopadhyay PK, and Roederer M (2004) Seventeen-colour flow cytometry: unraveling the immune system. Nat Rev Immunol 4:648–655 (http://www.ncbi.nlm.nih.gov/pubmed/15286731)) and more, but it takes a lot of planning and testing to make it work. The Attune NxT Acoustic Focusing Cytometer can run up to 14 colors plus forward and side scatter with a system equipped with all four lasers (violet, blue, yellow and red).
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
An FMO control (flow minus one) is a control in which you label cells or beads with every color in your panel, omitting one. Make one FMO control for each color in your panel. These controls are important for helping you properly set gates on your data.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The flow cytometry field is moving away from using isotype controls as they are not necessarily the most appropriate way to control for non-specific binding. Instead, they are using unstained cells to define the negative population, single stained controls to set compensation, and flow minus one (FMO) controls to set regions and gates.
You may want to consider whether using an isotype control is something you need to do. Here are some references you might want to look at:
-O'Gorman MR, Thomas J. (1999) Isotype controls-time to let go? Cytometry 38:78-80.
-Keeney M, Gratama JW, Chin-Yee IH et al. (1998) Isotype controls in the analysis of lymphocytes and CD34+ stem/progenitor cells by flow cytometry- time to let go! Cytometry 34:280-283.
-Hulspas R, O'Gorman MR, Wood BL et al. (2009) Considerations for the control of background fluorescence in clinical flow cytometry. Cytometry B Clin Cytom 76:355–364.
-Enumeration of Immunologically Defined Cell Populations by Flow Cytometry; Approved Guideline - Second Edition. Clinical and Laboratory Standards Institute, (CLSI). Document H42-A2 Volume 27 No.16, 2007.
-Clinical Flow Cytometric Analysis of Neoplastic Hematolymphoid Cells; Approved Guideline - Second Edition. Clinical and Laboratory Standards Institute, (CLSI). Document H43-A2 Volume 27 No. 11, 2007.
If you do wish to use isotype controls, we offer a wide variety of species and fluorophores, search our Web site under "isotype antibody control."
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Our Flow Cytometry Size Calibration Kit has non-fluorescent particle-size calibration standards that provide a simple, accurate way to determine cell sizes by flow cytometry. The kit contains six suspensions of highly uniform polystyrene microspheres with the following diameters: 1.0 µm, 2.0 µm, 4.0 µm, 6.0 µm, 10.0 µm and 15.0 µm. All of the microsphere suspensions are provided in convenient dropper vials. This kit looks at the scatter properties of non-fluorescent microspheres of known sizes to determine approximate cell sizes.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The smallest size that you can detect with the Attune NxT Acoustic Focusing Cytometer is 0.5 µm.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
The Attune NxT Autosampler, an optional accessory for the Attune NxT Acoustic Focusing Cytometer, enables rapid processing of up to 384 samples. It has broad compatibility with different plate formats, both 96- and 384-well plates. It has an intelligent probe designed to minimize clogging and carryover (<0.5%) and to prevent damage to the instrument. It mixes by aspiration rather than shaking to ensure homogeneity of the sample and maintain cell viability. Is performs automated cleaning as part of the shutdown process of the Attune NxT Cytometer. It provides consistent data regardless of sampling method (tube vs. plate) and collection rate.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
-Modular design - Multiple configurations available - field upgradable.
-Save time - 10X faster speeds with no loss in data quality.
-Simplified sample prep - No wash, no lyse options, non-clogging fluidics.
-Enables unique applications - Complex protocols on a broad range of cell types and samples.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
With the option to be configured with up to 4 lasers and 14 colors for multi-parameter analysis the Attune NxT Acoustic Focusing Cytometer was designed as a modular system to fit most experimental needs and lab budgets. The novel design of the optical path helps ensure precise fixed alignment of four spatially separated lasers onto the sample stream enabling consistency in data over time, superior performance, and superior reliability. The instrument can be configured with up to 4 solid-state lasers (405 nm, 488 nm, 561 nm, and 637 nm) with flat top beam profiles.
The Attune NxT Flow Cytometer's acoustic focusing uses ultrasonic radiation pressure (>
2 MHz) to transport particles into the center of the sample stream. This pre-focused stream is then injected into the sheath stream, which supplies an additional hydrodynamic pressure to the sample. The combination of these two forces- termed acoustic-assisted hydrodynamic focusing-results in a narrow core stream and uniform laser illumination, regardless of the sample input rate. In traditional cytometers that rely solely on hydrodynamic focusing, the sample core widens to accommodate the increases in flow rate, which results in less uniform laser light illumination.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.
Cytometry is the measurement of physical or chemical characteristics of cells or particles. Flow cytometry measures these characteristics of cells or particles as they individually pass lasers in a flow cytometer instrument. Flow cytometry is performed on single cells, providing discrete measurements for each cell in the sample. It also provides a statistical distribution of the measured characteristics of the sample.
A flow cytometer is made up of three subsystems: fluidics, optics, and electronics. Fluidics moves the cells and introduces them for interrogation. Optics generates and collects the light signals. Electronics converts the optical signals to proportional electronic signals for computer analysis.
Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.