| |
|
| |
Until now the utility of the violet laser has been limited by availability of compatible dyes. Moreover, organic fluorophores that are compatible
with the violet laser are often hindered by low signal to noise ratios and their requirement for extensive compensation.
Due to their intrinsic spectral advantages,
eFluor® Nanocrystals are brighter than commonly used
|
|
fluorochromes excited by the violet laser. The bright staining afforded by eFluor® Nanocrystals provides high signal to noise ratios and expands the
choices available when designing multicolor flow cytometry experiments. Look for our rapidly expanding portfolio of reagents to give you even greater flexibility and choice
with the violet laser.
|
| |
| |
| |
|
|
|
Comparison of eFluor® 605NC and Pacific Orange® staining. |
|
Mouse splenocytes were stained with Pacific Blue© anti-mouse CD3 (17A2) and eFluor® 605NC anti-mouse B220 (RA3-6B2) (left panel) or
Pacific OrangeŽ anti-mouse B220 (RA3-6B2) (right panel). Total viable cells in the lymphocyte gate were used for analysis. The data were collected using a violet laser octogan configured with the following filters: 450/50 BP (Pacific BlueŽ), 557 LP and 575/26 BP (Pacific OrangeŽ), 595 LP and 605/40 BP (eFluor 605NC).
|
|
| |
| |
Top
Flow Cytometry Products
Flow Cytometry Protocol
|
| |
| |
|
| |
With the flexible excitation wavelength of eFluor® Nanocrystals and their minimal compensation requirements, the combinations of dyes that can
be used in an experiment are vast. Procedurally, eFluor® Nanocrystals staining can be carried out in the same tube as organic dyes, and thus
|
|
eFluor® Nanocrystals can be easily integrated into any experiment. eBioscience offers a large and continually expanding selection of eFluor® Nanocrystals
directly-conjugated primary antibodies for flow cytometry.
|
| |
| |
|
| |
| |
eFluor® Nanocrystals are fully compatible with standard fluorochromes. |
|
| |
8-color phenotyping of human PBMC-derived Tregs was carried out using Alexa Fluor® 700 CD19 (HIB19), PerCP-Cy5.5 CD3 (OKT3), APC-Alexa Fluor® 750 CD8 (OKT8), eFluor® 605NC
|
|
CD4 (OKT4), APC CD127 (eBioRDR5), PE-Cy7 CD25 (BC96), FITC CD39 (A1) and PE CD101 (BB27). Total viable cells in the lymphocyte gate were used for analysis.
|
|
|
| |
| |
Top
Flow Cytometry Products
Flow Cytometry Protocol
|
| |
| |
|
| |
As part of the development process, we have evaluated the performance of eFluor® Nanocrystals(NC) in various staining buffers to determine optimal
staining conditions. Although eFluor® Nanocrystals staining is compatible
|
|
with eBioscience Flow Cytometry Staining Buffer, optimal staining is achieved when using eFluor® NC Flow Cytometry Staining Buffer.
|
| |
| |
|
|
|
Buffer comparison for eFluor® Nanocrystals staining. |
|
Mouse splenocytes were stained with eFluor® 605NC anti-mouse CD4 (GK1.5) in eBioscience
Flow Cytometry Staining Buffer (dashed line) or eFluor® NC Flow Cytometry Staining
Buffer (solid line). Viable cells in the lymphocyte gate were used for analysis.
|
|
| |
| |
Top
Flow Cytometry Products
Flow Cytometry Protocol
|
| |
| |
|
| |
Cells stained with eFluor® Nanocrystals can be fixed in paraformaldehyde with minimal loss of
fluorescent signal thereby allowing your samples to be analyzed at a later time providing you with
|
|
greater scheduling flexibility. We are currently evaluating the performance of eFluor®
Nanocrystals for use with intracellular staining protocols.
|
| |
| |
|
|
|
eFluor® Nanocrystals staining of live and fixed cells. |
|
Mouse splenocytes were stained with eFluor® 605NC anti-mouse B220 (RA3-6B2) and acquired as live cells (solid line) or after fixation
in 2% paraformaldehyde for 30 minutes at room temperature (dashed line). Cells in the lymphocyte gate were used for analysis.
|
|
| |
| |
Top
Flow Cytometry Products
Flow Cytometry Protocol
|
| |
| |
Overview of eFluor® Nanocrystals
Complete list of eFluor® Nanocrystals Products
|
| |
| |
|