CFP. PVR BV421 TX24 BD CD3 BUV496 UCHT1 BD CD8 BUV805 SK1 BD CD11b BV785 ICRF44 Biolegend CD14 APC-H7 MP9 BD CD16 BUV395 3G8 BD CD19 APC-Cy7 SJ25C1 . efluor 450. Fixable Violet DAPI SYTOX Blue Vybrant DyeCycle Violet FxCycle Violet Annexin V Pacific Blue PO-PRO-1 CellTrace Violet Click-iT Plus Edu Pacific Blue Azurite Cerulean eBFP eCFP mTurquoise Sirius BV421 eFluor 450 Horizon V450 VioBlue 512/25 VL2 Pacific Green LIVE/DEAD Fixable Aqua Violet Ratiometric Probe . the invitrogen live/dead violet viability/vitality kit provides a two-color fluorescence cell viability and vitality assay based on the simultaneous determination of live and dead cells with two probes that measure recognized parameters of cell health: plasma membrane integrity as a measure of cell viability, and intracellular esterase activity CD73 blocking antibody (anti-human CD73 Abs) and TLR-7 agonist were obtained from . D . LIVE/DEAD Fixable Dead Cell Stains are fixable viability dyes designed for accurate cell viability readouts in samples after fixation and/or permeabilization. User Changeable Fluorescence Filters Choose from six color options to run two-color assays Nave CD8 T cells were isolated from matched HNSCC or healthy donor PBMC using human nave CD8 T-cell isolating kit (STEMCELL Technologies) and then were stained with CellTrace Violet (5 mol/L, Life Technologies). : . Blue LASER 488 nm. For cytotoxicity assay, K562 or HT29 cells were labelled with CellTrace Violet (tested with BV421 laser channel, Invitrogen, USA) and then solely (negative control, NC) or co-cultured with NK cells derived from the indicated total mononuclear cells (MNCs) (UC-NKs, P-NKs) at a series of effector-to-target ratios (E: T). CellTrace CFSE . Violet 405 nm (30 mW) H 450/50 BV421 DAPI | Pacific Blue | BV450 |efluor450 | AF405 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet L/D Fix Violet | Zombie Violet | Sytox Blue . . cart()tcelltrace violetcd19+k562il-2fortessa x-50ctv Proliferated CADM1 + MHC-II + cells were considered as potential Flt3L-derived DCs. All-in-One System Basic cell counting, primary cell viability, and cell-based assays. To determine if the CD3xPDL1 BiTE kills PDL1+ tumor cells, PBMC from healthy donors were incubated at varying ratios with CellTrace Violet-labeled PDL1+ C8161 cells in the presence or absence of BiTE. Wash one time with 2 ml BioLegend's Cell Staining Buffer (Cat. DC - Vybrant DyeCycle (Thermo Fisher Scientific Inc.) BV xxx - BD Horizon Brilliant Violet (BD Biosciences Inc.) L/D/F . Superior performance bright, single-peak staining enables visualization of multiple generations Long-term signal stability well retained in cells for several days post-stain Its configuration is especially adapted to the use of new generation fluorochromes such as Brilliant Violet and Brilliant UltraViolet dyes. CellTrace calcein Violet AM, LIVE/DEAD Fixable Violet, DAPI, BV421 Indo-1 (blue) 505 LP 525/50 Indo-1 (blue), LIVE/DEAD Fixable Aqua Indo-1(violet) None 440/50 Alexa350, Indo-1(violet), CellTrace Calcein . Violet 405 nm (50 mW) VL1 472SP 445/45 BV421 DAPI | Pacific Blue | BV450 |efluor450 | AF405 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet L/D Fix Violet | Zombie Violet | Sytox Blue . Auckland Cytometry is a flow cytometry core facility which provides researchers with technical expertise, training and access to state-of-the-art technologies and applications in flow cytometry. 7. Violet 405 nm (50 mW) H 450/50 BV421 DAPI | Pacific Blue | BV450 |efluor450 | AF405 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet L/D Fix Violet | Zombie Violet | Sytox Blue . Unique Optical Collection and Analytical Capabilities. VIOLET V 450/50 Alexa Fluor 405, BD Horizon V450, BFP, BV421, Calcein Violet, Celltrace Violet, CFP, DAPI, DyLight 405, DyeCycle Violet, eFluor 450, Fixable Violet Dead Cell Stain, Fixable Viability eFluor 450, FxCycle Violet, Hoechst (blue,333258), Pacific Blue, PO-PRO-1, SYTOX Blue, VioBlue, V 525/50 CellTracker Violet BMQC is a fluorescent dye well suited for monitoring cell movement or location. When using two or more Super Bright, Brilliant Violet, Brilliant Ultra Violet, or other polymer dye- conjugated antibodies in a staining panel, we recommend . 420201) or equivalent buffer containing serum or BSA. PBMCs were labeled using the CellTrace Violet Cell Proliferation Kit (Invitrogen) according to the manufacturer's instructions. We can support your research by offering access to: training to use the analysers. Proliferation CFSE CellTrace Yellow CellTrace Violet CellTrace Far Red Cell Cycle - Live (Fisher) Vybrant Dye Cycle Green Vybrant Dye Cycle Ruby DNA Dyes/Cell Cycle (NOT Viability) Hoechst 33342/33258 : L34958 AF635 | AF647 | AF680 | Cy5 | Cy5.5| DyLight 633 CellTrace Far Red DyeCycle Ruby L/D Fix Far Red B 710LP 730/45 APC-Cy5.5 AF700 | APC-R700 Fixable Viability . The daughter cells of each subsequent generation inherit the dye allowing for long term analysis of proliferation. The violet excitation/emission spectra are ideal for multiplexing with green and red fluorescent dyes and proteins. BV421 BV605, Super Bright 600 N9-20017 Rev B Violet Laser Excited Dyes with Unique Signatures (2 of 2) 35 BV785, Super Bright 780 Qdot 800 N9-20017 Rev B Blue Laser Excited Dyes with Unique Signatures 43 cFluor B515, BB515, Vio 515, CellTrace Calcein Green AM, Live-or-Dye 488/515 cFluor B532, Alexa Fluor 488, CFSE, FITC, Migration assay analysis. Cy5 | Cy5.5| DyLight 633 CellTrace Far Red DyeCycle Ruby L/D Fix Far Red B 690LP 710/50 APC-CY5-5 AF700 | APC 650 | APC-R700 Fixable Viability 700 A . Superior performancebright, single-peak staining enables visualization of multiple generations Long-term signal stabilitywell-retained in cells for several days post stain BV421; SB436; Pacific Blue; Horizon V450; AF405; eFluor 450; CFP; DC Violet; SYTOX Blue; CellTrace calcein Violet AM; L/D/F Violet; DAPI 405C-525 505 LP 525/50 BV480; BV510; AmCyan; Pacific Orange; AF430; CFP; . BB515 BB700 PE PE-CF594 PE-Cy5 PE-Cy7 BV421 BV510 BV605 BV650 BV711 BV786 APC redFluor 710 APC-H7 BUV395 (379/28) BUV496 BUV661 . Violet 405 nm (100 mW) 450/50 BV421 Pacific Blue | BV450 | eFluor 450 | AF 405 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet DAPI | LIVE/DEAD Fixable Violet | Zombie Violet | Sytox Blue 525/50 BV510 BV480 | V500 | AF 430 | AmCyan | Pacific Green | Qdot 525 CFP | Cerulean LIVE/DEAD Fixable Aqua | Zombie Aqua . We incubate cells for 20 min at 37 degrees, and measure after 3/4 days. BV421 is excited by the 405 nm violet laser and emits at 421 nm. The y-axis shows median fluorescence intensity (MFI). Figure Legend Snippet: Normal trafficking of pmel-1 CD8 + T cells following 4-1BB triggering in vivo. CellTrace Violet CellTrace Far Red Cell Cycle (Fisher) Vybrant Dye Cycle Green Vybrant Dye Cycle Ruby Vybrant Dye Cycle Violet DNA Dyes/Cell Cycle (NOT Viability) Hoechst 33342/33258 . For this purpose, BMHCs were labeled with 5 M CellTrace Violet (Fisher Scientific, Spain), washed, and seeded at 0.8 10 6 cells/600 l into the 24-well plates. 6. Target cells were labelled with Cell Trace Violet (ThermoFisher) in PBS for 10 min at 37C, washed, then co-cultured with PBMCs in triplicate wells at the specified effector:target ratios. BUV661670/30 BUV737 740/35 BUV737 BUV805 820/60 BUV805 BV421 431/28 Brilliant Violet 421 | Horizon V450 Pacific Blue SuperBright 436 | Alexa 430 BFP CellTrace Violet DyeCycle Violet LIVE/DEAD Fixable Violet Zombie Violet KLRG1 expression and dilution of CellTrace-Violet in splenic CD45.1 + NK cells were analyzed 7 days after transfer. . Violet 406 nm (200 mW) 450/40 AF 405 DAPI | BV421 | Pacific Blue | BV450 | eFluor 450 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet DAPI | LIVE/DEAD Fixable Violet | Zombie Violet | Sytox Blue 515/20 AmCyan BV480 | BV510 | V500 | AF 430 | Pacific Green | Qdot 525 CFP | Cerulean LIVE/DEAD Fixable Aqua | Zombie Aqua . The x-axis shows each detector on the system organized first by laser line (UV, V, etc.) BB515 BB700 PE PE-CF594 PE-Cy5 PE-Cy5.5 PE-Cy7 BV421 BV510 BV605 BV650 BV786 APC redFluor 710 APC-H7 BUV395 BUV496 . Easily, reliably and quantitatively analyze live and dead cells within minutes The ability to stain live cells with a viability dye and preserve that staining pattern after fixation is critical for intracellular immunophenotyping. APC was sorted by FSC versus SSC in bulk. (A) Proliferative capacity of CD4 + and CD8 + T cells, determined by the percentage of CellTrace dim cells and the concentration of IFN- . Exclusion of the dead cells from the data allows cleaner separation and identification of cell populations. VIOLET V 450/50 Alexa Fluor 405, BD Horizon V450, BFP, BV421, Calcein Violet, Celltrace Violet, CFP, DAPI, DyLight 405, DyeCycle Violet, eFluor 450, Fixable Violet Dead Cell Stain, Fixable Viability eFluor 450, FxCycle Violet, Hoechst (blue, 333258), Pacific Blue, PO-PRO-1, SYTOX Blue, VioBlue V 525/50 AmCyan, BD Horizon V500, BV510. BV421 Rat Anti-Mouse CD40 (3/23) BD: Cat# 562846, RRID: AB_2734767: Brilliant Violet 510 anti-mouse I-A/I-E Antibody (M5/114.15.2) BioLegend: . with solid color bars beneath the chart. Proliferation CFSE CellTrace Yellow CellTrace Violet CellTrace Far Red CellTrace Blue (379/28) Cell Cycle - Live (Fisher) Vybrant Dye Cycle Green Vybrant Dye Cycle Ruby Vybrant Dye Cycle Violet DNA Dyes/Cell Cycle Created Date: Page 3 Rev. Figure S6 continued. Briefly, PBMCs were stained with CellTrace Violet at a concentration of 1.0 10 6 cells/mL for 20 min at 37C. Violet FxCycle Violet Annexin V, Pacific Blue Annexin V, eFluor 450 PO-PRO-1 CellTrace Violet Click-iT Plus EdU Pacific Blue Azurite Cerulean TagBFP ECFP mTurquoise AmCyan BD Horizon BV421 BD Horizon V450 VioBlue 510/50 520/35 525/50 530/40 550/40 VL2 512/25 VL2(V6) 525/50 eFluor 506 Pacific Green LIVE/DEAD Fixable Aqua Violet Ratiometric Probe . 300 x g 5 OpTmizer T SFM . the presence of BiTE and tumor, whereas only 27% proliferated in the presence of BiTE, and 1.4% in the absence of BiTE and tumor. No. antiCD3 PE, antiCD4 BV421, and CD8 APC antibodies. CFSE and other similar dyes (CellTrace Violet, FarRed etc) cross the cellular membrane in living cells and bind covalently and permanently to intracellular structures (usually lysine or other amines). Invitrogen Pacific Blue CellTrace Violet FxCycle VioletLIVE/DEAD Fixable Violet BV421 eFluor 450 Invitrogen Pacific Green F2N12S AmCyan BV510 Permanently label cells with Invitrogen CellTrace fluorescent stains without affecting morphology or physiology to trace generations or divisions in vivo or in vitro. Cells can be fixed with paraformaldehyde or methanol prior to permeabilization or can be analyzed without fixation. Violet (16 channels) Blue (14 channels) Red (8 channels) Yellow-Green (10 channels) This dye is excited by all 5 lasers. We then added 1% FBS in PBS (Corning) to the cells for 5 min at RT to quench unbound dye. To assess the functional potential of Tim-3 + and Tim-3 + Tregs or Tim-3 Violet CellTrace-stained iDCs (K) . Cy5 | Cy5.5| DyLight 633 CellTrace Far Red DyeCycle Ruby L/D Fix Far Red B 690LP 710/50 APC-CY5.5 AF700 | APC-R700 Fixable Viability 700 A . Cy5 | Cy5.5 CellTrace Far Red DyeCycle Ruby LIVE/DEAD Fixable Far Red FL11 720/60 AF 700 APC-Cy5.5 | APC-R700 Fixable Viability 700 FL12 785/60 APC-AF750 . (B) CellTrace-Violet-labeled WT NK cells were transferred into WT mice (CD45.2 +), Rag1 / mice (CD45.2 +), or Rag1 / Nod2 / mice (CD45.2 +). Incubate the cells at room temperature, in the dark, for 15-30 minutes. CellTrace Violet followed by activation with antiCD3 antibody. Proliferation was assessed at 0, 1.5, 3, 6, 9, 12, and 14 days of incubation by flow cytometry. Tables 1,2,3. . Cells were then analyzed on the Agilent NovoCyte Quanteon for cell proliferation of both the CD4 and Dual-Fluorescence for Accurate Primary Cell Viability No interference from red blood cells. Proliferation CFSE CellTrace Yellow CellTrace Violet CellTrace Far Red CellTrace Blue Cell Cycle - Live (Fisher) Vybrant Dye Cycle Green Vybrant Dye Cycle Ruby Vybrant Dye Cycle Violet DNA Dyes/Cell Cycle (NOT Viability) OT-I nuclei were identified as CellTrace Violet hi CD3e low. (i.e. Configuration of the instruments is detailed in the . C57BL/6 mice were injected intravenously with CFSE-labeled pmel-1 Thy1.1 + CD8 + T cells ( a , c , d ) or CellTrace Violet-labeled S1PR1-GFP pmel-1 Thy1.1 + CD8 + T cells ( b ), immunized with hgp100 peptide in IFA, and further stimulated with rat IgG or anti-4-1BB mAb at days 0 and 2. This dye has a medium-to-high relative brightness and can be used for cell surface, intracellular, and nuclear staining. CellTrace CFSE Cell Proliferation Kit is used for in vitro and in vivo labeling of cells to trace multiple generations using dye dilution by flow cytometry. (D) Histograms (left) show c-Fos staining of isolated nuclei from control (black histograms) or PTBP1-deficient (blue histograms) nave CD8 T cells cocultured with bulk splenocytes pulsed with 10 nM of OVA peptide at the time indicated. CellTrace Violet Cell Proliferation Kit, for flow cytometry: Molecular Probes: Cat# C34557: Critical commercial assays: Chromium Single Cell 30 reagent kits v1 . Pacific Blue CellTrace Violet FxCycle VioletLIVE/DEAD Fixable Violet BV421 eFluor 450 : Pacific Green F2N12S AmCyan BV510 : : . The T cell proliferation assay in PBMCs was performed as follows: 1. 14 Aug 2022 Analyzers LSR Fortessa Analyzer BD Biosciences Laser Ex (nm) Em (nm) Channel Fluorochromes Ultra-violet 355 379/28 355-B BUV395 Ultra-violet 355 450/50 355-A DAPI, L/D blue, Zombie UV, Hoechst 33342 Violet 405 450/50 405-F Pacific Blue, BV421, eFluor450, SB436 Violet 405 525/50 405-E L/D Aqua, BV510, AmCyan Violet 405 610/20 405-D BV605, SB600 . CellTrace violet-labeled mononuclear cells from DLBCL (n = 13) and control group (n = 12 [rLN, n = 3; cHL, n = 6; FL, n = 3]) tissues were activated by plate-coated CD3 and soluble CD28 antibodies. BB515 BB700 PE PE-CF594 PE-Cy5 PE-Cy5.5 PE-Cy7 BV421 BV510 BV605 APC redFluor 710 APC-H7 SuperBright 600 . 40 mL OpTmizer T SFM . 5. PBMCs were isolated using . trained staff to assist with analysis or cell sorting. : 20 . 37C 20 . Spectral flow cytometry (SFC) is based on many of the fundamental aspects of conventional flow cytometry but has unique optical collection and analytical capabilities. Data are representative of three independent experiments with 3 mice in . Cy5 | Cy5.5 | DyLight 633 CellTrace Far Red DyeCycle Ruby L/D Fix Far Red RL2 735LP 780/60 APC-CY7 APC-H7 | APC-Fire750 | APC-eFluor 780 | AF750 . The ratio of CD8 + plus CD4 + T cells to tumor cells is 2.3:1. 10 mL CellTrace Violet . (104)-PE +, CD62L(MEL-14)-BV421 +, CD44(IM7 Analyze bone marrow, peripheral blood, and cord blood without lysing. We are trying this with Celltrace violet. CellTrace Violet. PBMC from a NSCLC patient were stained for MDSC. 405 BV421 450/50 BV421, Pacific Blue, VioBlue, Alexa405, Calcien Blue FVS450*, SYTOX Blue, Zombie Violet* DAPI VPD450, CellTrace Violet, DyeCycle Violet 405 BV510 525/50 BV510, Pacific Green, AmCyan FVS510*, Zombie Aqua* 405 BV605 610/20 BV605, Pacific Orange Zombie Yellow* 488 FITC 530/30 5 . Violet 404 nm (200 mW) H 410LP 431/28 BV421 DAPI | Pacific Blue | BV450 |efluor450 | AF405 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet L/D Fix Violet | Zombie Violet | Sytox Blue . We tried different concentrations, 2 uM, 5 uM and 20 uM for 200.000 - 500.000 cells. . you can't use BV421 with DAPI), but is brightest in the 355B channel, so take your measurement there. Cells were stained with 7AAD and the CellTrace violet stained tumor cells were gated and analyzed for % dead cells. Where indicated, T cells were stained before co-culture with CellTrace Violet (Invitrogen) for proliferation assays, as per manufacturer protocol. Violet 402 nm (200 mW) H 410LP 431/28 BV421 Pacific Blue | BV450 |efluor450 | AF405 | VioBlue | DyLight 405 BFP | Azurite CellTrace Violet DyeCycle Violet L/D Fix Violet | Zombie Violet | Sytox Blue . Violet 405 nm 445/45 VL1 Horizon BV421 Pacific Blue Alexa Fluor 405 eBFP Azurite Cerulean Calcein Violet SYTOX Blue LIVE/DEAD Fixable Violet DAPI CellTrace Violet Click-iT Plus EdU- Pacific Blue DAPI FXCycle Violet Vybrant DyeCycle Violet 530/30 VL2 Pacific Green Horizon BV510 VioGreen vGFP LIVE/DEAD Fixable Aqua 572/28 VL3 Horizon BV570 . BV421; SB436; Pacific Blue; Horizon V450; AF405; eFluor 450; CFP; DC Violet; SYTOX Blue; CellTrace calcein Violet AM; L/D/F Violet; DAPI VL2 495 DLP 512/25 BV480; BV510; AmCyan; eFluor 506; Pacific Green; AF430; L/D/F Aqua VL3 555 DLP 603/48 BV570; BV605; SB600; eFluor 605NC; Pacific Orange; L/D/F Yellow Data are pooled from two independent experiments with PBMC from two individual patients. GD2-iCAR PBT persistence and immune phenotype in patients.E) Patient 101, F) Patient 201 and G) Patient 203. i) CD8+ T-cell immune phenotype, and ii) CD4+ and CD8+ T cell PD-1/PD-L1 expression profile for Patient 201 who received FluCy-mediated lymphodepleting chemotherapy on days -4, -3 and -2 before BV421. For co-culture, a total of 100,000 T cells and roughly 20 enteroids/well were co-cultured in 200l per well of complete culture medium with 30% Matrigel in 24-well plates. AAVS1-19bbz cells were labelled with CellTrace Violet (Thermo Fisher) according to the manufacturer's instructions. Raji cells were pretreated with mitomycin C (50 g ml -1 ) for 90 min at 37 . *BV, BUV, BB dyes require the use of the Brilliant . With SFC, the emission spectrum of every fluorescent molecule is captured by a set of detectors or an array of channels . DAPI450/50 CellTrace Blue Hoechst 33342 LIVE/DEAD Fixable Blue | Zombie UV BUV496515/30 BUV563586/15 . Superior performancebright, single-peak staining enables visualization of multiple generations Long-term signal stabilitywell-retained in cells for several days post stain The CellTrace Violet and CellTracker Violet/Green flow assay kits were obtained from Life Technologies (USA). A range of popular division tracking dyes were subsequently developed, including CellTrace Violet (CTV), making available the green fluorescent channel previously occupied by CFSE. Violet 405 nm (10 mW) FL6 450/50 BV421 Pacific Blue | BV450 | eFluor 450 | AF 405 | VioBlue BFP | Azurite CellTrace Violet DyeCycle Violet DAPI | LIVE/DEAD Fixable Violet | Zombie Violet . AF635 | AF647 | AF680 | Cy5 | Cy5.5| DyLight 633 CellTrace Far Red DyeCycle Ruby L/D Fix Far Red B 710LP 730/45 APC-Cy5.5 AF700 | APC-R700 Fixable Viability . (5 min/500g) and stained using CellTrace Violet BV421 (Thermo Fisher Scientific) for 15 min at 37 C, and then washed with PBS . Our cell viability dyes for flow cytometry are designed to measure cell membrane integrity or intracellular esterase activity. eFluor 450, DyeCycle Violet, SYTOX Blue, CellTrace calcein Violet AM, LIVE/DEAD Fixable Violet, DAPI, BV421 APC-Cy7 735 LP 780/60 APC-Alexa750, APC-Cy7, APC-eFluor780 APC None 660/20 Alexa647, APC, SYTOX Red, DRAQ5 Blue Violet Red CantoII Filter Configuration. COVID-19 Update: Learn about our efforts to help battle coronavirus. CellTrace Violet Cell Proliferation Kit is used for in vitro and in vivo labeling of cells to trace multiple generations using dye dilution by flow cytometry. Our affordable spectral flow cytometry instruments make complex applications possible. After loading into cells, the dye is well retained, allowing for multigenerational tracking of cellular movements. Violet (405) UV (355nm) Red (640nm) Yellow/Green (561nm) LSRII Filter Configuration. CellTrace Violet Click-iT Plus EdU Paci c Blue Azurite Cerulean TagBFP ECFP mTurquoise AmCyan BD Horizon BV421 eFluor 450 BD Horizon V450 VioBlue 510/50 512/25 520/35 525/50 530/40 542/50 550/40 VL2 512/25 Paci c Green LIVE/DEAD Fixable Aqua Violet Ratiometric Probe (F2N12S) T-Sapphire BD Horizon BV510 BD Horizon . 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