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Molecular Probes™ Flow Cytometry Sub-micron Particle Size Reference Kit

Catalog No. F13839
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F13839 6 x 2 mL
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Catalog No. F13839 Supplier Molecular Probes™ Supplier No. F13839
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Description

Flow Cytometry Sub-micron Particle Size Reference Kit

The Flow Cytometry Sub-micron Particle Size Reference Kit provides six sizes of green-fluorescent beads (ranging in diameter from 0.02μm to 2.0μm) packaged in individual vials. Each bead is uniform in size, and the green-fluorescent dyes are localized within the bead, which makes the beads brighter and more photostable than conventional surface-stained beads.

These size standards can help identify small bioparticles such as exosomes, bacteria, and viruses in experimental samples. This kit can also be used as a tool to verify instrument performance and establish parameters suitable for analyzing sub-micron particles, including:
  • Particle size resolution limit and dynamic range
  • Sensitivity of forward and side scatter photomultiplier tubes
  • Level of instrument baseline noise
  • Laser and optical alignment and stability
  • Stability of the fluidics system

    The bead diameters are determined by transmission electron microscopy, and the beads are provided as 2 mL suspensions with a density of ∼1× 106 beads/mL.

    • Specifications

    Diameter (Metric) 0.02, 0.1, 0.2, 0.5, 1.0, 2.0 μm
    Content And Storage Store at 2°C to 8°C. DO NOT FREEZE.
    Quantity 6 x 2 mL
    Color Green
    Product Type Flow Cytometry Particle Size Reference Kit

    Frequently Asked Questions (FAQs)

    I am using the Flow Cytometry Sub-micron Particle Size Reference Kit and can't resolve the 0.1 and 0.2 µm beads in my flow run. What can I do to get good separation?

    We recommend mixing the beads thoroughly before use and using a slower flow rate.

    Are there any surface modifications of the beads within the Flow Cytometry Sub-micron Particle Size Reference Kit?

    Yes. Charged groups are on the surface making the beads water-dispersible.
    Where is the dye localized in the beads within the Flow Cytometry Sub-micron Particle Size Reference Kit?

    The dye is dispersed throughout the polystyrene matrix.

    When using the Live/Dead BacLight Bacterial Viability and Counting Kit, for flow cytometry, some cells seem to have both red and green signal. Are these cells dead or alive or dying?

    The green dye in the kit will label all the cells as it is a cell-permeant nucleic acid stain. The red dye is not cell permeant, and will only stain the cells with compromised membranes (dead cells). Therefore, any cells with red signal will be considered dead. It is possible that you will have some cells that are only red, some that are red and green, and some that are only green. Sometimes the red dye will displace the green dye. In any case, any red cells are dead.

    Also, the green dye emission may bleed through into the red channel. Do a single-color staining and examine under both green and red filter sets to determine the level of bleedthrough. To avoid this bleedthrough, use a lower concentration of dye, and, if possible, use narrow bandpass filters.
    How can I count my bacteria by flow cytometry?

    There are several options. We have two fluorescence based kits that are useful for bacterial counting: Live/Dead BacLight Bacterial Viability and Counting Kit, for flow cytometry (Cat. No. L34856) and Bacteria Counting kit, for flow cytometry (Cat. No. B7277). Another option is the Flow Cytometry Sub-micron Particle Size Reference Kit (Cat. No. F13839).
    What is the smallest size that I can detect with the Attune NxT Acoustic Focusing Cytometer?

    The smallest size that you can detect with the Attune NxT Acoustic Focusing Cytometer is 0.5 µm.
    What is the Attune NxT Autosampler?

    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.
    What are the advantages of acoustic-assisted hydrodynamic focusing in flow cytometry?

    -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.
    How is the Attune NxT Acoustic Focusing Cytometer different from traditional flow cytometers?

    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.
    What is flow cytometry?

    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.

    For Research Use Only. Not for use in diagnostic procedures.