多色流式細胞儀



Violet Laser Overview

螢光物質

 

BD Horizon™ BV421

BD Horizon™ Brilliant Violet™ 421 (BV421) (Ex-Max 407 nm/Em-Max 421 nm) is a polymer-based dye excited by the violet laser and is one of the brightest fluorochromes offered by BD Biosciences. Conjugates are typically 10 times brighter than Pacific Blue™ conjugates and are often as bright as or brighter than PE conjugates. Due to nearly identical excitation and emission properties but different spillover characteristics, BV421, Pacific Blue™, and BD Horizon™ V450 cannot be used simultaneously.

Filter: 450/40
Relative brightness: Brightest (on a scale from Dim, Moderate, Bright, and Brightest)
Sample data:

Comparison data from two regulatory T-cell panels
Comparison data from two regulatory T-cell panels

Figures 3a and 3b compare the brightness of CD25 BV421 with CD25 PE. Figures 3c and 3d compare the brightness of CD127 BV421 with CD127 Alexa Fluor® 647. Figures 3a–3d: Data is shown on CD3+ lymphocytes.

Figures 3e and 3f show how the BV421 reagents contribute to the unequivocal identification of the regulatory T-cell population. Figures 3e–3f: Data is shown on CD3+CD4+ lymphocytes.

T-cell subset resolved: CD8 memory and effector T cells
T-cell subset resolved: CD8 memory and effector T cells
T-cell subset resolved: CD4 memory and effector T cells
T-cell subset resolved: CD4 memory and effector T cells
T-cell subset resolved: Regulatory T cells
T-cell subset resolved: Regulatory T cells

7 TNK 方案的資料舉例

7 TNK 方案的資料舉

例如:在 7 色方案中使用 CD8 Brilliant Violet 421 ,識別 T 細胞和 NK 細胞。裂解的全血用 CD8 Brilliant Violet 421 CD3 APC-H7 CD7 PE CD16 PE-Cy™7 CD45 BD Horizon V500 CD56 APC 以及 CD57 FITC 染色。在 BD FACSVerse 流式細胞儀上檢測。

Example of BV421 used in an intracellular staining protocol
Example of BV421 used in an intracellular staining protocol

Peripheral blood mononuclear cells (PBMCs) stimulated with staphylococcus enterotoxin B (SEB) for 6 hours in the presence of brefeldin A (BFA). Cells were then washed, fixed with BD FACS™ lysing solution, permeabilized with BD FACS™ permeabilizing solution 2, and stained with CD8 BV421, CD3 FITC, CD4 PerCP-Cy™5.5, IFN-γ PE, and IL-2 APC. The panel was run on a BD FACSVerse flow cytometer and data is shown on CD3+ gated lymphocytes.

Example of BV421 staining using various protocols
Example of BV421 staining using various protocols
Histogram overlays of human CD3 staining using various staining protocols: live-cell stain, cell staining post-fixation/permeabilization, or cell staining prior to fixation/permeabilization.
Analysis of human T-cell subsets
Analysis of human T cell subsets
Data shows a 10-color panel run on a 3-laser BD LSRFortessa™ X-20 with a blue/red/violet configuration. The panel was used for defining various subsets of regulatory T cells (A, B), memory/effector CD4+ T cells (C, D, E, F), and memory/effector CD8+ T cells (G, H, I, J).
Various subsets of regulatory T cells
Various subsets of regulatory T cells
Memory/effector CD4+ and CD8+ T cells
Memory/effector CD4+ and CD8+ T cells

 
 

BD Horizon™ V450

BD Horizon V450 (Ex-Max 404 nm/Em-Max 448 nm) is a coumarin dye excited by the violet laser that exhibits spectral properties similar to Pacific Blue™. Conjugates are typically as bright or brighter than comparable reagents conjugated to Pacific Blue™. Due to nearly identical excitation and emission properties but different spillover characteristics, BV421, Pacific Blue™, and BD Horizon V450 cannot be used simultaneously.

Filter: 450/40
Relative brightness: Dim (on a scale from Dim, Moderate, Bright, and Brightest)

 

BD Horizon™ V500

BD Horizon V500 (Ex-Max 415 nm/Em-Max 500 nm) is a novel organic dye excited by the violet laser. This dye offers improved brightness over Pacific Orange™ and reduced spillover into the FITC channel when compared to AmCyan. BD Horizon V500 cannot be used simultaneously with AmCyan or Pacific Orange™.

Filter: 525/50
Relative brightness: Dim (on a scale from Dim, Moderate, Bright, and Brightest)

CD4 BD Horizon V500 comparison to Pacific Orange™
CD4 BD Horizon V500 comparison to Pacific Orange™
Freshly isolated human lymphocytes stained with CD4 (RPA-T4) conjugated to BD Horizon V500, Pacific Orange™, or AmCyan. Samples run on a BD FACSCanto™ II flow cytometer. BD Horizon V500 and AmCyan data was collected with a 510/50-nm, 502-LP filter. Pacific Orange™ data was collected with a 585/42-nm filter. The plots have been superimposed to show signal comparison.
Analysis of human lymphocytes: T-cell subsets
Analysis of human lymphocytes: T-cell subsets
Data shows a 10-color panel run on a 3-laser BD LSRFortessa X-20 with a blue/red/violet configuration. The panel was used for defining various lymphocyte T-cell subsets (A–E) and non–T cell subsets (F–J) using lysed whole blood (BD FACS lysing solution) from a normal donor.
Analysis of human lymphocytes: Non–T-cell subsets
Analysis of human lymphocytes: Non–T-cell subsets
Data shows a 10-color panel run on a 3-laser BD LSRFortessa X-20 with a blue/red/violet configuration. The panel was used for defining various lymphocyte T-cell subsets (A–E) and non–T-cell subsets (F–J) using lysed whole blood (BD FACS lysing solution) from a normal donor.

 
 

BD Horizon™ BV510

BD Horizon Brilliant™ Violet 510 (BV510) (Ex-Max 405 nm/Em-Max 510 nm) is a polymer-based dye. BD Horizon BV510 can be excited by the violet laser and detected in the BD Horizon V500 (525/50-nm) filter set. Due to similar excitation and emission properties, BD Horizon BV510 and BD Horizon V500 cannot be used simultaneously.

Filter: 525/50
Relative brightness: Moderate (on a scale from Dim, Moderate, Bright, and Brightest)
Sample data:

Example data for CD11c BV510 and CD19 BV510
Example data for CD11c BV510 and CD19 BV510
Panel A shows BALB/c mouse splenocytes stained with mouse CD11c BD Horizon BV510 (black line) or an isotype control (red dashed line). Panel B shows lysed whole blood stained with human CD19 BD Horizon BV510. Data shown was gated on lymphocytes and overlaid with the isotype control.
Stain index comparison between BD Horizon BV510 and FITC
Stain index comparison between BD Horizon BV510 and FITC
BD Horizon BV510 and FITC reagents of the same clone were run to compare the stain index.
 

BD Horizon™ BV605

BD Horizon Brilliant™ Violet 605 (BV605) (Ex-Max 407 nm/Em-Max 602 nm) is a tandem fluorochrome that combines BD Horizon BV421 and an acceptor dye. These conjugates are very bright, exhibiting brightness similar to equivalent PE conjugates. Due to the excitation of the acceptor dye by the green (532-nm) and yellow-green (561-nm) lasers, there will be significant spillover of the BV605 signal into the PE and BD Horizon™ PE-CF594 detectors off the green or yellow-green lasers.

Filter: 610/20
Relative brightness: Bright (on a scale from Dim, Moderate, Bright, and Brightest)
Sample data:

Example data for BV605.
Figure 2. Example data for BV605.
Lysed whole blood stained with CD19 or CD4 BV605. Data shown was gated on lymphocytes.
Example data from a human hematopoietic stem cell panel
Example data from a human hematopoietic stem cell panel
Frozen human cord blood was thawed and then lineage-depleted. The lineage-depleted cord blood was then stained with FITC (CD2, CD3, CD4, CD7, CD8, CD10, CD11b, CD14, CD19, CD20, CD56, and CD235a), BV421 CD38, BV605 CD90, APC CD34, APC-H7 CD45RA, and 7-AAD.
Example of BV605 staining using various protocols
Example of BV605 staining using various protocols
Histogram overlays of human CD4 BV605 staining using various staining protocols: live-cell stain, cell staining prior to fixation/permeabilization, or cell staining post-fixation/permeabilization.

 
 

BD Horizon™ BV711

BD Horizon Brilliant™ Violet 711 (BV711) (Ex-Max 407 nm/Em-Max 711 nm) is a tandem fluorochrome of BD Horizon BV421 and an acceptor dye with an Em Max at 711 nm. This dye offers a very bright choice for the violet laser. Due to the excitation and emission characteristics of the acceptor dye, there may be moderate spillover into the Alexa Fluor® 700 and PerCP-Cy™5.5 detectors.

Filter: 710/50
Relative brightness: Brightest (on a scale from Dim, Moderate, Bright, and Brightest)
Sample data:

Example data for CD335 (NKp46) BV711 and CD19 BV711
Example data for CD335 (NKp46) BV711 and CD19 BV711
Lysed whole blood stained with human CD335 (NKp46) BV711 (A) or CD19 BV711 (B). Data shown was gated on lymphocytes and overlaid with the isotype control.
Various subsets of memory/effector CD4+ and CD8+ T cells
Various subsets of memory/effector CD4+ and CD8+ T cells
BD Horizon Brilliant Violet dyes detecting various subsets of CD8+ T cells
BD Horizon Brilliant Violet dyes detecting various subsets of CD8+ T cells
BD Horizon Brilliant Violet dyes detecting various subsets of memory/effector CD4+ T cells
BD Horizon Brilliant Violet dyes detecting various subsets of memory/effector CD4+ T cells


 


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

Alexa Fluor® is a registered trademark, Pacific Orange™ and Pacific Blue™ are trademarks of Life Technologies Corporation.

Cy™ is a trademark of GE Healthcare. Cy™ dyes are subject to proprietary rights of GE Healthcare and Carnegie Mellon University and are made and sold under license from GE Healthcare only for research and in vitro diagnostic use. Any other use requires a commercial sublicense from GE Healthcare, 800 Centennial Avenue, Piscataway, NJ 08855-1327, USA.

CF™ is a trademark of Biotium, Inc.