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Celltrace violet fluorescent dye

Manufactured by Thermo Fisher Scientific
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CellTrace Violet is a fluorescent dye used for labeling and tracking cells. It binds to cellular proteins, allowing for the identification and monitoring of cell populations. The dye has an excitation maximum at 405 nm and an emission maximum at 450 nm, making it compatible with common flow cytometry and fluorescence microscopy setups.

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Lab products found in correlation

Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (2 mentions) Celltrace cell proliferation kit, by Thermo Fisher Scientific (1 mentions) Buv805, by BD (1 mentions) Facscelesta flow cytometer, by BD (1 mentions)

Close spelling variants of this product

CellTrace Violet dye CellTrace Violet CellTrace dye Fluorescent dyes

5 protocols using celltrace violet fluorescent dye

1

Evaluating T-cell Responses to SpCas9 Epitopes

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Splenocyte clearance assays were performed similarly to previous work96. Briefly, spleens from adult C57BL/6J mice were harvested and treated to remove erythrocytes and dead cells. These cells were then diluted to 107 cells/ml and split into two pools, one of which was pulsed for 40 min with a pool of the 30 most immunogenic T-cell epitopes in SpCas9 (according to our predictions) at 1 μg/ml each and labeled with the CellTrace Violet fluorescent dye (ThermoFisher). The other pool was pulsed with a matching amount of DMSO, and labeled with the green fluorescent dye CFSE (ThermoFisher). A 1:1 mixture of these cells were then injected into naïve or CFA-immunized mice at week 1 or 3.5 retro-orbitally at 3–6 × 107 cells per mouse. Spleens from these mice were harvested 16–20 hours later, treated to remove erythrocytes, and analyzed by flow cytometry to assess the degree of specific clearance of the CTV+ cells which were pulsed with Cas9 peptides.
Moreno A.M., Palmer N., Alemán F., Chen G., Pla A., Jiang N., Chew W.L., Law M, & Mali P. (2019). immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy. Nature biomedical engineering, 3(10), 806-816.
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2

Labeling T. cruzi Trypomastigotes with Fluorescent Dyes

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Cell suspensions of T. cruzi trypomastigotes were labelled with CellTrace Violet fluorescent dye (CellTrace Cell Proliferation Kit, ThermoFisher Scientific, Waltham, MA) and CFSE following manufacturer’s instructions. Briefly, 2 × 106 trypomastigotes were incubated for 20 min at 37°C with 10 μM of CellTrace Violet or for 5 min with 5 μM of CFSE, protected from light. Unbound dye was quenched by the addition of five volumes of 10% FBS-RPMI or one volume of FBS respectively. After washing in fresh media parasites were used for infection of cell cultures or mice.
Sánchez-Valdéz F.J., Padilla A., Wang W., Orr D, & Tarleton R.L. (2018). Spontaneous dormancy protects Trypanosoma cruzi during extended drug exposure. eLife, 7, e34039.
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3

Profiling PBMC T-cell Responses

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Participant PBMCs were thawed using a CryoThaw adaptor48 (link) into 10 mL of RPMI supplemented with 10% FBS. PBMCs were rested for approximately 4 h following thaw. Subsequently, 1e6 cells were labeled with cell trace violet fluorescent dye following the manufacturer’s instructions (Thermo Fisher) and cultured in 1 mL of RPMI supplemented with 10% FBS in FACS tubes and stimulated for five days with 1 mg/mL ancestral peptides (JPT Peptide Technologies) that have been resuspended in DMSO. Unstimulated samples were supplemented with equivalent volume DMSO. After five days, surface staining was performed for flow cytometry using Invitrogen Live/Dead Fixable Blue Dead Cell Stain and anti-human CD3 (BUV661, BD Biosciences), CD4 (BUV805, BD Biosciences), CD8α (BV510, BioLegend), CCR7 (BUV395, BD Biosciences) and CD45RA (APC-H7, BD Biosciences) antibodies. T-cell proliferation assays were performed on participants for which we had sufficient PBMC samples.
Roznik K., Xue J., Stavrakis G., Johnston T.S., Kalluri D., Ohsie R., Qin C.X., McAteer J., Segev D.L., Mogul D., Werbel W.A., Karaba A.H., Thompson E.A, & Cox A.L. (2024). COVID-19 vaccination induces distinct T-cell responses in pediatric solid organ transplant recipients and immunocompetent children. NPJ Vaccines, 9, 73.
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4

MIA PaCa-2 Cell Growth Kinetics

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MIA PaCa-2 cells were pre-cultured at 5 % CO2, Continuous 10 % CO2 or intermittent exposure to 10 % CO2 (3 hrs/day). Cells were labelled with CellTrace™ Violet fluorescent dye (Thermo Fisher Scientific, Waltham, MA‎, USA) as per manufacturer recommendations and left to grow in their respective conditions for 5 days. Cells were then collected and assessed using a BD FACS Celesta™ Flow cytometer (Becton, Dickinson and Company [BD], Warwick, Rhode Island, USA) using an excitation/emission of 405nm/450nm. Data were analysed using the FlowJO software (FlowJo LLC, Ashland, OR, USA).
Nevler A., Brown S.Z., Nauheim D., Portocarrero C., Bassig J., Schultz C.W., McCarthy G., Lavu H., Yeo T.P., Yeo C.J, & Brody J.R. (2020). Effects of Hypercapnia, an Element of Obstructive Respiratory Disorders on Pancreatic Cancer Chemoresistance and Progression. Journal of the American College of Surgeons, 230(4), 659-667.
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5

Evaluating T-cell Responses to SpCas9 Epitopes

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Splenocyte clearance assays were performed similarly to previous work96. Briefly, spleens from adult C57BL/6J mice were harvested and treated to remove erythrocytes and dead cells. These cells were then diluted to 107 cells/ml and split into two pools, one of which was pulsed for 40 min with a pool of the 30 most immunogenic T-cell epitopes in SpCas9 (according to our predictions) at 1 μg/ml each and labeled with the CellTrace Violet fluorescent dye (ThermoFisher). The other pool was pulsed with a matching amount of DMSO, and labeled with the green fluorescent dye CFSE (ThermoFisher). A 1:1 mixture of these cells were then injected into naïve or CFA-immunized mice at week 1 or 3.5 retro-orbitally at 3–6 × 107 cells per mouse. Spleens from these mice were harvested 16–20 hours later, treated to remove erythrocytes, and analyzed by flow cytometry to assess the degree of specific clearance of the CTV+ cells which were pulsed with Cas9 peptides.
Moreno A.M., Palmer N., Alemán F., Chen G., Pla A., Jiang N., Chew W.L., Law M, & Mali P. (2019). immune-orthogonal orthologues of AAV capsids and of Cas9 circumvent the immune response to the administration of gene therapy. Nature biomedical engineering, 3(10), 806-816.
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