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View additional product information for AbC™ Total Antibody Compensation Bead Kit - FAQs (A10513, A10497)
24 product FAQs found
这是可能的。但是如果您的值超过100，可能需要查看你们用来采集数据的电压。
通过运行单色对照，有可能除去溢出到另一个荧光采集通道的荧光信号。
进行补偿时，你需要为自己使用的每种颜色参数准备单一染色样品（或补偿微球）。此外，我们建议您使用FMO（荧光扣除）对照。您可以使用减去一种颜色的其他所有颜色标记您的细胞或者微球来作为对照。为各种颜色准备FMO对照。这些对照对正确设置数据门限很有帮助。
理想情况下，每一个单独荧光团的荧光发射光谱都非常集中，窄峰，发射峰之间分隔的较开。现实中，有机染料和荧光蛋白有很宽的发射峰，必须考虑补偿（数据采集中或后）来矫正任何一个荧光团荧光信号的分配。补偿从重叠光谱中移除了荧光信号，使您知道所看见的信号就是目标荧光团发出的信号，所以非常重要。
补偿对照仅需未染色的细胞样品（或阴性对照）和单一颜色标记的样品——每管一个。如果您希望单色染色对照如您期望的那么亮,，那么可以使用我们的AbC总抗体补偿微珠试剂盒（货号A10513和A10497），这试剂盒是将抗体结合到微珠而非细胞上。
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。
Check the isotype of the two antibodies to see if they are different. If so, we offer isotype-specific goat anti-mouse secondary antibodies that can be used together with minimal cross-reactivity. If the two mouse antibodies are of the same isotype, then you would need to make direct conjugates, such as with our antibody labeling kits or APEX, and SiteClick kits.
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Here are some tips to follow:
- Make sure that the beads were never frozen.
- Avoid mixing by vortexing too long (no longer than 10 seconds) or sonication as this may promote denaturation of the protein on the surface of the beads.
- Use the product within the warranty period (one year).
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Yes, the AbC Total Antibody Compensation Bead Kit can bind Fab dimers as long as the Fab dimers are derived from either mouse, rat, hamster, or rabbit species.
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As an alternative, you may covalently attach your antibody to colorless, 5 µm Aldehyde/Sulfate Latex Beads (Cat. No. A37306). The aldehyde moiety on the bead surface directly binds with primary amines on the surface of the antibodies in simple buffers such as PBS (pH 7.4). Make sure that the buffer does not contain any primary amines (i.e., do not use Tris or glycine buffers). To avoid making bead-antibody-bead-antibody multimers, add excess antibody relative to the beads.
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It is possible. But if you have a lot of values over 100, you probably need to look at the voltages that you are using for your data collection.
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By running single-color controls, it is possible to remove signal of one fluorophore that spills over into the collection channel for another fluorophore.
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For compensation, you need to prepare a singly stained sample (or compensation beads) for each color parameter that you are using. In addition, we recommend that you use FMO (flow minus one) controls. These are controls in which you label cells or beads with every color in your panel, omitting one. Make one FMO control for each color. These controls are important for helping you properly set gates on your data.
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In a perfect world, the fluorescence emission profile for each individual fluorophore would be a very intense, narrow peak, well separated from all other emission peaks. In reality, organic dyes and fluorescent proteins have broad emission peaks, and compensation must be employed (during or after data acquisition) to correctly assign fluorescence signal to each fluorophore. Compensation is important because it removes fluorescent signal from overlapping spectra so you know that the signal you see is only the signal from the fluorophore of interest.
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All that you need for compensation controls is an unstained sample of your cells (or negative control beads) and samples with each of your fluorophores - one per tube. You want the single-stained controls to be as bright as you expect your brightest sample to be. Antibodies can be bound to beads instead of cells using our AbC Total Antibody Compensation Bead Kit (Cat. Nos. A10513 and A10497).
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The smallest size that you can detect with the Attune NxT Acoustic Focusing Cytometer is 0.5 µm.
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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.
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-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.
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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.
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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.
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