time lapse experiments z stacks Search Results


a1r confocal microscope  (Nikon)
99
Nikon a1r confocal microscope
A1r Confocal Microscope, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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deltavision omx sr imaging system  (Cytiva Europe)
96
Cytiva Europe deltavision omx sr imaging system
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Deltavision Omx Sr Imaging System, supplied by Cytiva Europe, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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gopro hero5 camera  (GoPro Inc)
90
GoPro Inc gopro hero5 camera
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Gopro Hero5 Camera, supplied by GoPro Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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caspase inhibitor z-vad-fmk g7231  (Promega)
90
Promega caspase inhibitor z-vad-fmk g7231
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Caspase Inhibitor Z Vad Fmk G7231, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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statgraphics centurion v17.1.02  (statpoint inc)
90
statpoint inc statgraphics centurion v17.1.02
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Statgraphics Centurion V17.1.02, supplied by statpoint inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/time+lapse+experiments+z+stacks/10__15446_slash_rcciquifa__v45n1__58026-49-24-38?v=statpoint+inc
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massspectrometry-based experiments  (DeGussa Corporation)
90
DeGussa Corporation massspectrometry-based experiments
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Massspectrometry Based Experiments, supplied by DeGussa Corporation, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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mid-infrared nanoimaging experiments  (NanoImaging Services Inc)
90
NanoImaging Services Inc mid-infrared nanoimaging experiments
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Mid Infrared Nanoimaging Experiments, supplied by NanoImaging Services Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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transcelerate estimand template  (TransCelerate Biopharma Inc)
90
TransCelerate Biopharma Inc transcelerate estimand template
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Transcelerate Estimand Template, supplied by TransCelerate Biopharma Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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experimental-chemie  (Chemie GmbH)
90
Chemie GmbH experimental-chemie
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Experimental Chemie, supplied by Chemie GmbH, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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bert-as-service  (TenCent Inc)
90
TenCent Inc bert-as-service
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Bert As Service, supplied by TenCent Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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rangen shrimp feed  (Rangen Inc)
90
Rangen Inc rangen shrimp feed
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Rangen Shrimp Feed, supplied by Rangen Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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experimental general experimental 1h  (Varian Medical)
86
Varian Medical experimental general experimental 1h
CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by <t>DeltaVision</t> workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.
Experimental General Experimental 1h, supplied by Varian Medical, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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experimental general experimental 1h - by Bioz Stars, 2026-08
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Image Search Results


CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by DeltaVision workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.

Journal: Cells

Article Title: Overcoming Immunological Resistance Enhances the Efficacy of a Novel Anti-tMUC1-CAR T Cell Treatment against Pancreatic Ductal Adenocarcinoma

doi: 10.3390/cells8091070

Figure Lengend Snippet: CAR architecture, expression on engineered T cells, and binding of CAR-T cells to target cells. ( A ) The architecture of three different CAR constructs used in this study. In the original construct (tMUC1-CAR), scFv of TAB004 Ab is linked to CD28 transmembrane (TM) domain followed by CD28 and CD3ζ intracellular domains in a retroviral plasmid. CD8a leader sequence was used as signal peptide for cell membrane expression of the CAR. In the CTL-CAR construct, scFv of TAB was removed. In the CAR-mKate construct, mKate2 gene was fused to the C-terminus of CAR flanking with a GA linker. ( B ) CTL-CAR and tMUC1-CAR expression measured by flow cytometry using FITC-conjugated anti-myc tag Ab, in CD4+ and CD8+ primary T cells on day 12 after infection. On average, 42% of T cells expressed tMUC1-CAR. ( C ) Bright field (top left) and fluorescent image (top right) of live T cells expressing CAR-mKate plated in 35 mm poly-D-lysine coated MatTek dish and imaged by DeltaVision workstation (Applied Precision, GE), projection image of a T cell expressing CAR-mKate (bottom left), and one Z image of the CAR-mKate T cell (bottom right) illustrating the ring-like structure around the cells formed by CAR-mkate expression, which indicates even distribution of CAR molecules on the T cell membrane. ( D ) Light and fluorescent image of CAR-mKate T cells binding to MUC1 expressing cancer cell (HPAFII). HPAFII cells were incubated with CAR-mKate T cells for 4 h, then T cells were removed, HPAFII cells were washed and imaged using DeltaVision microscope. The intense red signal observed between CAR T cell and HPAFII indicates co-localization and binding of CAR molecules, which suggests the formation of immunological synapse. Nuclei were stained with Hoechst nuclei blue dye in C and D. All scale bars = 15 μm.

Article Snippet: Videos of CAR T cells killing BxPC3-MUC1 vs. BxPC3-Neo cells were taken by time lapse imaging using a DeltaVision OMX-SR imaging system (#29115476, GE).

Techniques: Expressing, Binding Assay, Construct, Plasmid Preparation, Sequencing, Flow Cytometry, Infection, Incubation, Microscopy, Staining