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spectral ami amix system  (Spectral Instruments Imaging)
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Spectral Instruments Imaging spectral ami amix system
Spectral Ami Amix System, supplied by Spectral Instruments Imaging, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/spectral+ami/ami+ht+ivis+spectral+system/pmc13128220-66-6-9
Average 86 stars, based on 1 article reviews
spectral ami amix system - by Bioz Stars, 2026-09
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spectral ami optical imaging system  (Spectral Instruments Imaging)
86
Spectral Instruments Imaging spectral ami optical imaging system
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Spectral Ami Optical Imaging System, supplied by Spectral Instruments Imaging, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/spectral+ami/ami+htx/pmc12002645-113-9-14
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spectral ami optical imaging system - by Bioz Stars, 2026-09
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spectral ami ht ivis system  (Spectral Instruments Imaging)
86
Spectral Instruments Imaging spectral ami ht ivis system
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Spectral Ami Ht Ivis System, supplied by Spectral Instruments Imaging, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/spectral+ami/ami+ht+ivis+spectral+system/pmc13200424-192-22-26
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spectral ami ht ivis system - by Bioz Stars, 2026-09
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spectral ami x optical imaging platform  (Spectral Instruments Imaging)
86
Spectral Instruments Imaging spectral ami x optical imaging platform
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Spectral Ami X Optical Imaging Platform, supplied by Spectral Instruments Imaging, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/spectral+ami/ami+htx/pm41088753-326-7-13
Average 86 stars, based on 1 article reviews
spectral ami x optical imaging platform - by Bioz Stars, 2026-09
86/100 stars
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ami spectral instruments imaging version 4.0.8  (Spectral Instruments Imaging)
90
Spectral Instruments Imaging ami spectral instruments imaging version 4.0.8
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Ami Spectral Instruments Imaging Version 4.0.8, supplied by Spectral Instruments Imaging, 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/spectral+ami/ami+spectral+instruments+imaging+version+4+0+8/pm40580639-570-15-16
Average 90 stars, based on 1 article reviews
ami spectral instruments imaging version 4.0.8 - by Bioz Stars, 2026-09
90/100 stars
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spectral ami-htx imaging  (HTX Technologies LLC)
90
HTX Technologies LLC spectral ami-htx imaging
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Spectral Ami Htx Imaging, supplied by HTX Technologies LLC, 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/spectral+ami/spectral+ami+htx+imaging+system/pm40473600-516-0-1
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spectral ami-htx imaging - by Bioz Stars, 2026-09
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spectral ami-htx imaging system  (HTX Technologies LLC)
90
HTX Technologies LLC spectral ami-htx imaging system
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Spectral Ami Htx Imaging System, supplied by HTX Technologies LLC, 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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spectral ami optical imaging system  (Bruker Corporation)
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Bruker Corporation spectral ami optical imaging system
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Spectral Ami Optical Imaging System, supplied by Bruker 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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ami-ht (spectral instruments imaging) imaging system  (Spectral Instruments Imaging)
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Spectral Instruments Imaging ami-ht (spectral instruments imaging) imaging system
(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the <t>Spectral</t> <t>Ami</t> <t>Optical</t> <t>Imaging</t> <t>System</t> over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.
Ami Ht (Spectral Instruments Imaging) Imaging System, supplied by Spectral Instruments Imaging, 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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(a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the Spectral Ami Optical Imaging System over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.

Journal: Journal of controlled release : official journal of the Controlled Release Society

Article Title: Timely administration of drug combination improves chemoimmunotherapy of an immune-cold tumor

doi: 10.1016/j.jconrel.2025.02.075

Figure Lengend Snippet: (a) Schematic depiction of 2E’/PTX/CDN assembly. (b) TEM image of 2E’/PTX/CDN (5:1:0.1, w/w/w). Scale bar: 100 nm. (c) In vitro PTX release kinetics from 2E’/PTX/CDN in PBS containing 0.2% Tween-80 (n = 3 replicates of a representative batch, mean ± SD). (d) Confocal images of bone marrow-derived dendritic cells (BMDCs) incubated with free CDN or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration equivalent to 0.1 µg/ml FITC-labeled CDN for 8 h. Cell membrane was stained with Wheat Germ Agglutinin Alexa Fluor 633 Conjugate. Red: cell membrane; green: CDN. Scale bar: 50 µm. (e) Fluorescence intensity of BMDCs incubated with PBS, free CDN, or 2E’/PTX/CDN (5:1:0.1, w/w/w) at a concentration of 0.1 µg/ml FITC-labeled CDN for 8 h, measured by flow cytometry. n=3 replicates of a representative batch. (f) TLR5 activation indicated by SEAP production from HEK-Blue ™ mTLR5 reporter cells incubated with 2E’ or 2E’/PTX/CDN (5:1:0.1, w/w/w) for 24 h. Flagellin (Fla, 100 ng/mL), a known TLR5 agonist, was used as a positive control, shown in blue. n=3 replicates in a representative experiment. (g) B16F10 cells undergoing apoptosis and necrosis, stained with Annexin V-PI. The cells were treated with PBS (negative control) or 2E’/PTX/CDN at 4 μg/mL 2E’, 0.8 μg/mL PTX, and 0.08 μg/mL CDN for 24 h. The fractions of cell population undergoing apoptosis and necrosis were determined by dual-color analysis; n = 3 replicates in a representative experiment. (h) Calreticulin (CRT) exposure and ATP secretion of B16F10 melanoma cells after 24 h incubation with 2E’ (1.25 µg/mL), PTX (0.25 µg/mL), 2E’/PTX (1.25 µg/mL 2E’; 0.25 µg/mL PTX), 2E’/siPD-L1 (1.25 µg/mL 2E’; 1.125 µg/mL siPD-L1), or 2E’/PTX/CDN (1.25 µg/mL 2E’; 0.25 µg/mL PTX; 0.025 µg/mL CDN). CRT exposure was measured by flow cytometry. See Fig. S5 for gating strategy. n=3 replicates in a representative experiment. (i) Maturation of BMDCs incubated with PBS, 2E’ (5 µg/mL), PTX (1 µg/mL), CDN (0.1 µg/mL), 2E’/PTX (5 µg/mL 2E’; 1 µg/mL PTX), 2E’/CDN (5 µg/mL 2E’; 0.1 µg/mL CDN), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN) for 24 h. n=3 replicates of a representative batch. (j) Left: Secretion of TNF-α and IFN-β by BMDCs incubated with PBS, 2E’ (5 µg/mL), CDN (0.1 µg/mL), or 2E’/PTX/CDN (5 µg/mL 2E’; 1 µg/mL PTX; 0.1 µg/mL CDN); right: TNF-α secretion by bone marrow-derived macrophages (BMDMs) incubated with free CDN or 2E’/PTX/CDN at 0.05 µg/mL CDN equivalent (CDN L and 2E’/PTX/CDN L ) or 0.1 µg/mL CDN equivalent (CDN H and 2E’/PTX/CDN H ). For both 2E’/PTX/CDN L and 2E’/PTX/CDN H , the weight ratio of 2E’:PTX:CDN was 5:1:0.1. n=3 replicates of a representative batch. (k) Cytotoxicity of 2E’/PTX/CDN to immune cells (BMDC and BMDM) and tumor cells (4T1 breast cancer cells, CT26 colon cancer cells, and B16F10 melanoma cells). The weight ratio of 2E’:PTX:CDN was 5:1:0.1 at all concentrations. n=2–3 replicates of a representative batch. (l) Fluorescence intensity of Cy7-CDN indicating tumor retention of CDN. Free CDN* or 2E’/PTX/CDN* (5:1:0.1, w/w/w) was intratumorally injected into CT26 subcutaneous tumor at a total dose of 0.02 mg CDN* (CDN + Cy7-labeled CDN in 1:1 (w/w) ratio). The intensity of Cy7-CDN fluorescence signal was monitored by the Spectral Ami Optical Imaging System over 7 d from the injection and expressed as values normalized to the intensity at the time 0, n=2 for free CDN; n=3 for 2E’/PTX/CDN, mean ± SD. An extended data set with the full timescale and replicates is shown in Fig. S7 . P values in d-f were calculated by Tukey’s multiple comparisons test following ordinary one-way ANOVA.

Article Snippet: The fluorescence signal of CDN was monitored by the Spectral Ami Optical Imaging System (Spectral Instruments, Tucson, AZ) over 7 d from the injection.

Techniques: In Vitro, Derivative Assay, Incubation, Concentration Assay, Labeling, Membrane, Staining, Fluorescence, Flow Cytometry, Activation Assay, Positive Control, Negative Control, Injection, Optical Imaging