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A Illustration of the ribosome profiling in phagocytes, macrophages, and cancer cells. Peripheral blood monocytes were isolated from healthy donors and differentiated into macrophages. The macrophages were labeled with CellTracker Blue and co-cultured with <t>CFSE-labeled</t> cancer cells for 2 h. Subsequently, flow cytometry was employed to sort the macrophages, phagocytes, and cancer cells as indicated. Cells were then subjected to ribosome profiling analysis. A total of 100 healthy donors were enrolled. B The volcano plot illustrates the identification of dysregulated genes, with phagocyte-specific genes determined by comparing phagocytes to MDA-MB-231 cancer cells (FPKM = 0 in MDA-MB-231 cells). Dysregulated genes were subsequently identified by comparing these phagocyte-specific genes to those in control macrophages. Differentially expressed genes (DEGs) were defined as those meeting the criteria of p < 0.05 and fold change > 2. FC fold change, FDR false detecting rate. C GO analysis of the dysregulated genes, dysregulated genes were identified, and Gene Ontology Analysis was applied to analyze the enriched pathways of dysregulated genes. D The detail of the dysregulated genes enriched pathways, gene ontology analysis was applied to analyze the dys-enriched pathways, the detail of the pathway, including the q value and the number of dysregulated genes, was shown. E The rank of the dysregulated receptor and transmembrane protein, PD1, Siglec-10, and CD37, was indicated, X -axis, the rank of the genes, Y -axis, the log 2 FC, FC fold change. F The rank of the dysregulated receptor and transmembrane protein (RPKM > 0 in phagocytes), X -axis, the rank of the genes, Y -axis, the log 2 FC, FC fold change.

Celltrace Cfse C34554, supplied by Thermo Fisher, 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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1) Product Images from "Targeting CD37 promotes macrophage-dependent phagocytosis of multiple cancer cell types and facilitates tumor clearance in mice"

Article Title: Targeting CD37 promotes macrophage-dependent phagocytosis of multiple cancer cell types and facilitates tumor clearance in mice

Journal: Nature Communications

doi: 10.1038/s41467-025-61348-2

Figure Legend Snippet: A Illustration of the ribosome profiling in phagocytes, macrophages, and cancer cells. Peripheral blood monocytes were isolated from healthy donors and differentiated into macrophages. The macrophages were labeled with CellTracker Blue and co-cultured with CFSE-labeled cancer cells for 2 h. Subsequently, flow cytometry was employed to sort the macrophages, phagocytes, and cancer cells as indicated. Cells were then subjected to ribosome profiling analysis. A total of 100 healthy donors were enrolled. B The volcano plot illustrates the identification of dysregulated genes, with phagocyte-specific genes determined by comparing phagocytes to MDA-MB-231 cancer cells (FPKM = 0 in MDA-MB-231 cells). Dysregulated genes were subsequently identified by comparing these phagocyte-specific genes to those in control macrophages. Differentially expressed genes (DEGs) were defined as those meeting the criteria of p < 0.05 and fold change > 2. FC fold change, FDR false detecting rate. C GO analysis of the dysregulated genes, dysregulated genes were identified, and Gene Ontology Analysis was applied to analyze the enriched pathways of dysregulated genes. D The detail of the dysregulated genes enriched pathways, gene ontology analysis was applied to analyze the dys-enriched pathways, the detail of the pathway, including the q value and the number of dysregulated genes, was shown. E The rank of the dysregulated receptor and transmembrane protein, PD1, Siglec-10, and CD37, was indicated, X -axis, the rank of the genes, Y -axis, the log 2 FC, FC fold change. F The rank of the dysregulated receptor and transmembrane protein (RPKM > 0 in phagocytes), X -axis, the rank of the genes, Y -axis, the log 2 FC, FC fold change.

Techniques Used: Isolation, Labeling, Cell Culture, Flow Cytometry, Control

Labeling:

Article Title: Optimization of hypo-alloimmunogenic multispecific CAR-T and SARS-CoV-2-specific T cells for off-the-shelf adoptive cell therapy
Article Snippet: .. The stimulator and responder cells were resuspended in PBS and labeled by 1 μM of CellTrace Violet (cat. C34557) and 1 μM of Celltrace CFSE (cat. C34554) (both from Thermo Fisher Scientific), respectively, at 37°C for 30 min. ..

Article Title: Boosting tumor homing of endogenous natural killer cells via therapeutic secretomes of chemically primed natural killer cells
Article Snippet: .. NK cells (5×10 5 ) were labeled with 1 μM CellTrace CFSE (C34554; Thermo Fisher Scientific) and loaded into the upper chamber in serum-free medium. ..

Article Title: Targeting CD37 promotes macrophage-dependent phagocytosis of multiple cancer cell types and facilitates tumor clearance in mice
Article Snippet: Macrophage was induced by PMA (100 ng/ml) (MedChemExpress, Catalog # HY-18739). .. Macrophages were then labeled with CellTracker CMF2HC (Thermo Fisher Scientific, Catalog # C12881 ), and breast cancer cells MDA-MB-231 were labeled with CellTrace CFSE (Thermo Fisher Scientific, Catalog # C34554 ). ..

Article Title: Mouse enteric neurons control intestinal plasmacytoid dendritic cell function via serotonin-HTR7 signaling
Article Snippet: .. To generate mature BMDCs, 1 μg/ml lipopolysaccharide (LPS; LPS25, Sigma) was added to the cell suspensions on day 8 for 24 h. At day 9, mature BMDCs were harvested, and labeled with CellTrace CFSE (C34554, Thermo Fisher Scientific) and then injected intravenously into Tph2 fl/fl and Tph2 fl/fl ; Hand2-Cre mice. ..

Article Title: TRBC1-CAR T cell therapy in peripheral T cell lymphoma: a phase 1/2 trial.
Article Snippet: .. Target cells were labeled with CellTrace CFSE (C34554, Thermo Fisher Scientific) following the manufacturer’s instructions. .. Mock and CAR-transduced T cells were labeled with CellTrace Violet (C34557, Thermo Fisher Scientific) following the manufacturer’s instructions.

Article Title: Boosting tumor homing of endogenous natural killer cells via therapeutic secretomes of chemically primed natural killer cells.
Article Snippet: .. NK cells (5×105) were labeled with 1 μM CellTrace CFSE (C34554; Thermo Fisher Scientific) and loaded into the upper chamber in serum- free medium. ..

Multiple Displacement Amplification:

Isolation:

Article Title: Low-Dose Cyclophosphamide Enhances the Tumoricidal Effects of 5-Day Spacing Stereotactic Ablative Radiotherapy by Boosting Antitumor Immunity
Article Snippet: .. The isolated CD8 + T cells were stained with 5 μM CellTrace CFSE (# C34554 , Thermo Fisher Scientific), stimulated with Dynabeads Mouse T Activator CD3/CD28 (#11452D, Gibco) at a 1:1 cell-to-bead ratio in a U-bottom 96-well plate (1.0×10 5 cells/200 μL/well), and treated with serial dilutions of Cy, starting from 6.4 mM. ..

Staining:

Injection:

other:

Article Title: Neoantigen-based mRNA vaccine exhibits superior anti-tumor activity compared to synthetic long peptides in an in vivo lung carcinoma model.
Article Snippet: For DiR staining, DiR (D12731, Thermo Fisher Scientific, USA) was added to the LNP at a concentration of 1% before mixing with mRNA and injected intravenously into the mice.

Image Search Results

Journal: Nature Communications

Article Title: Targeting CD37 promotes macrophage-dependent phagocytosis of multiple cancer cell types and facilitates tumor clearance in mice

doi: 10.1038/s41467-025-61348-2

Figure Lengend Snippet: A Illustration of the ribosome profiling in phagocytes, macrophages, and cancer cells. Peripheral blood monocytes were isolated from healthy donors and differentiated into macrophages. The macrophages were labeled with CellTracker Blue and co-cultured with CFSE-labeled cancer cells for 2 h. Subsequently, flow cytometry was employed to sort the macrophages, phagocytes, and cancer cells as indicated. Cells were then subjected to ribosome profiling analysis. A total of 100 healthy donors were enrolled. B The volcano plot illustrates the identification of dysregulated genes, with phagocyte-specific genes determined by comparing phagocytes to MDA-MB-231 cancer cells (FPKM = 0 in MDA-MB-231 cells). Dysregulated genes were subsequently identified by comparing these phagocyte-specific genes to those in control macrophages. Differentially expressed genes (DEGs) were defined as those meeting the criteria of p < 0.05 and fold change > 2. FC fold change, FDR false detecting rate. C GO analysis of the dysregulated genes, dysregulated genes were identified, and Gene Ontology Analysis was applied to analyze the enriched pathways of dysregulated genes. D The detail of the dysregulated genes enriched pathways, gene ontology analysis was applied to analyze the dys-enriched pathways, the detail of the pathway, including the q value and the number of dysregulated genes, was shown. E The rank of the dysregulated receptor and transmembrane protein, PD1, Siglec-10, and CD37, was indicated, X -axis, the rank of the genes, Y -axis, the log 2 FC, FC fold change. F The rank of the dysregulated receptor and transmembrane protein (RPKM > 0 in phagocytes), X -axis, the rank of the genes, Y -axis, the log 2 FC, FC fold change.

Article Snippet: Macrophages were then labeled with CellTracker CMF2HC (Thermo Fisher Scientific, Catalog # C12881 ), and breast cancer cells MDA-MB-231 were labeled with CellTrace CFSE (Thermo Fisher Scientific, Catalog # C34554 ).

Techniques: Isolation, Labeling, Cell Culture, Flow Cytometry, Control

EFE-184 and AN3CA cells were administered two CRISPR/Cas9 lentivirus vectors (ARNTL-KO 1, 2#). mRNA ( A ) and protein ( B ) expression of ARNTL in WT or ARNTL-KO EC cells examined by RT-qPCR and WB analysis, respectively; C the proliferation of EC cells analyzed by CCK-8 method; D apoptosis and necrosis of EC cells analyzed by YO-PRO-1/PI staining; migration ( E ) and invasion ( F ) of EC cells determined by Transwell assays; G the proportion of M2 macrophages after co-culture with WT or ARNTL-KO EC cells analyzed by flow cytometry; mRNA ( H ) and protein ( I ) expression of M1 phenotype markers (IL-6 and TNF-α) and M2 markers (PDL-1 and IL-10) in the co-cultured macrophages analyzed by RT-qPCR and WB analysis, respectively; J proliferation of CD8 + T cells co-cultured with WT or ARNTL-KO EC cells analyzed by CFSE labeling; K concentrations of secretory IFN-γ in the co-culture system of EC cells and CD8 + T cells analyzed by ELISA. All cellular experiments were performed in five biological replicates. Differences were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Journal: Cell Death & Disease

Article Title: ARNTL-mediated INO80-DHX15 axis reprograms the glycolytic metabolism and augments the progression of endometrial carcinoma

doi: 10.1038/s41419-025-07776-w

Figure Lengend Snippet: EFE-184 and AN3CA cells were administered two CRISPR/Cas9 lentivirus vectors (ARNTL-KO 1, 2#). mRNA ( A ) and protein ( B ) expression of ARNTL in WT or ARNTL-KO EC cells examined by RT-qPCR and WB analysis, respectively; C the proliferation of EC cells analyzed by CCK-8 method; D apoptosis and necrosis of EC cells analyzed by YO-PRO-1/PI staining; migration ( E ) and invasion ( F ) of EC cells determined by Transwell assays; G the proportion of M2 macrophages after co-culture with WT or ARNTL-KO EC cells analyzed by flow cytometry; mRNA ( H ) and protein ( I ) expression of M1 phenotype markers (IL-6 and TNF-α) and M2 markers (PDL-1 and IL-10) in the co-cultured macrophages analyzed by RT-qPCR and WB analysis, respectively; J proliferation of CD8 + T cells co-cultured with WT or ARNTL-KO EC cells analyzed by CFSE labeling; K concentrations of secretory IFN-γ in the co-culture system of EC cells and CD8 + T cells analyzed by ELISA. All cellular experiments were performed in five biological replicates. Differences were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test. ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: According to the instructions of the CellTrace carboxyfluorescein diacetate succinimidyl ester (CFSE) Cell Proliferation Kit ( C34554 , Thermo Fisher Scientific), the CD8 + T cells were incubated with CFSE solution (1 mM) at 37 °C for 15 min for labeling, followed by incubation with EC cells.

Techniques: CRISPR, Expressing, Quantitative RT-PCR, CCK-8 Assay, Staining, Migration, Co-Culture Assay, Flow Cytometry, Cell Culture, Labeling, Enzyme-linked Immunosorbent Assay

ARNTL-KO EC cells were further administered Vector-INO80 or Vector NC. A mRNA expression of INO80 in EC cells determined by RT-qPCR; B intracellular INO80, Bcl-2, Bax, Cleaved Caspase-3 protein expression by WB analysis; C DNA replication of EC cells analyzed by EdU labeling assay; D apoptosis and necrosis of EC cells analyzed by YO-PRO-1/PI staining; migration ( E ) and invasion ( F ) of EC cells determined by Transwell assays; G the concentrations of TGF-β1, PGE2, and IL-10 released from EC cells and the intracellular concentration of IDO by ELISA; H proportion of M2 macrophages after co-cultured with the treated EC cells analyzed by flow cytometry; I expression of M1 phenotype markers (IL-6 and TNF-α) and M2 markers (PDL-1 and IL-10) in the co-cultured macrophages analyzed by RT-qPCR; J proliferation of CD8 + T cells co-cultured with the EC cells analyzed by CFSE labeling; K concentrations of secretory IFN-γ in the co-culture system of EC cells and CD8 + T cells analyzed by ELISA. All cellular experiments were performed in five biological replicates. Differences were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test. **** p < 0.0001.

Journal: Cell Death & Disease

Article Title: ARNTL-mediated INO80-DHX15 axis reprograms the glycolytic metabolism and augments the progression of endometrial carcinoma

doi: 10.1038/s41419-025-07776-w

Figure Lengend Snippet: ARNTL-KO EC cells were further administered Vector-INO80 or Vector NC. A mRNA expression of INO80 in EC cells determined by RT-qPCR; B intracellular INO80, Bcl-2, Bax, Cleaved Caspase-3 protein expression by WB analysis; C DNA replication of EC cells analyzed by EdU labeling assay; D apoptosis and necrosis of EC cells analyzed by YO-PRO-1/PI staining; migration ( E ) and invasion ( F ) of EC cells determined by Transwell assays; G the concentrations of TGF-β1, PGE2, and IL-10 released from EC cells and the intracellular concentration of IDO by ELISA; H proportion of M2 macrophages after co-cultured with the treated EC cells analyzed by flow cytometry; I expression of M1 phenotype markers (IL-6 and TNF-α) and M2 markers (PDL-1 and IL-10) in the co-cultured macrophages analyzed by RT-qPCR; J proliferation of CD8 + T cells co-cultured with the EC cells analyzed by CFSE labeling; K concentrations of secretory IFN-γ in the co-culture system of EC cells and CD8 + T cells analyzed by ELISA. All cellular experiments were performed in five biological replicates. Differences were analyzed by two-way ANOVA followed by Sidak’s multiple comparisons test. **** p < 0.0001.

Techniques: Plasmid Preparation, Expressing, Quantitative RT-PCR, Labeling, Staining, Migration, Concentration Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Flow Cytometry, Co-Culture Assay

Vector DHX15/Vector NC was administered into the ARNTL-KO EC cells. A The proportion of M2 macrophages after co-cultured with the EC cells analyzed by flow cytometry ( n = 3); B proliferation of CD8 + T cells co-cultured with the EC cells analyzed by CFSE labeling ( n = 3); C proliferation of the EC cells analyzed by CCK-8 method ( n = 3). AN3CA cells administered ARNTL-KO 1 + Vector-INO80/Vector NC were injected into immunocompetent huHSC-NOG-EXL mice subcutaneously. D volume of xenograft tumors in mice ( n = 10); E weight of xenograft tumors on day 28 and the representative images ( n = 10); F lactate content in xenograft tumors analyzed by colorimetry ( n = 5); G number of infiltrating CD8 + T cells and macrophages in xenograft tumors determined by flow cytometry ( n = 5); migration ( H ) and invasion ( I ) of EC cells determined by Transwell assays ( n = 3). AN3CA cells administered ARNTL-KO 1 + Vector-INO80/Vector NC were injected into immunocompetent huHSC-NOG-EXL mice via the tail vein. J survival of mice within 45 days after tail vein injection of EC cells ( n = 10); K tumor infiltration in mouse lung tissues analyzed by HE staining ( n = 10). Differences were analyzed by the unpaired t test ( E , F , K ) or two-way ANOVA followed by Sidak’s multiple comparisons tests ( A – D , G – I ). All cellular experiments were performed in three biological replicates. Survival of mice was analyzed by the Log-rank test ( J ). *** p < 0.001, **** p < 0.0001.

Journal: Cell Death & Disease

Article Title: ARNTL-mediated INO80-DHX15 axis reprograms the glycolytic metabolism and augments the progression of endometrial carcinoma

doi: 10.1038/s41419-025-07776-w

Figure Lengend Snippet: Vector DHX15/Vector NC was administered into the ARNTL-KO EC cells. A The proportion of M2 macrophages after co-cultured with the EC cells analyzed by flow cytometry ( n = 3); B proliferation of CD8 + T cells co-cultured with the EC cells analyzed by CFSE labeling ( n = 3); C proliferation of the EC cells analyzed by CCK-8 method ( n = 3). AN3CA cells administered ARNTL-KO 1 + Vector-INO80/Vector NC were injected into immunocompetent huHSC-NOG-EXL mice subcutaneously. D volume of xenograft tumors in mice ( n = 10); E weight of xenograft tumors on day 28 and the representative images ( n = 10); F lactate content in xenograft tumors analyzed by colorimetry ( n = 5); G number of infiltrating CD8 + T cells and macrophages in xenograft tumors determined by flow cytometry ( n = 5); migration ( H ) and invasion ( I ) of EC cells determined by Transwell assays ( n = 3). AN3CA cells administered ARNTL-KO 1 + Vector-INO80/Vector NC were injected into immunocompetent huHSC-NOG-EXL mice via the tail vein. J survival of mice within 45 days after tail vein injection of EC cells ( n = 10); K tumor infiltration in mouse lung tissues analyzed by HE staining ( n = 10). Differences were analyzed by the unpaired t test ( E , F , K ) or two-way ANOVA followed by Sidak’s multiple comparisons tests ( A – D , G – I ). All cellular experiments were performed in three biological replicates. Survival of mice was analyzed by the Log-rank test ( J ). *** p < 0.001, **** p < 0.0001.

Techniques: Plasmid Preparation, Cell Culture, Flow Cytometry, Labeling, CCK-8 Assay, Injection, Colorimetric Assay, Migration, Staining

(A) Groups of OVA-immunized or naïve C57Bl/6 mice ( n =9/group) were injected intradermally in the ear at time zero with 2 μg OVA and 5 μg each of adjuvants polyI:C and pam3Cys. 60 hrs later, sampling microneedle were applied to the same site for 12 hrs, followed by retrieval and flow cytometry analysis. (B) Sample processing and flow cytometry gating strategy. (C-D) Representative flow cytometry plots ( C ) and quantification from groups of animals ( D ) showing OVA-specific SIINFEKL/H-2K b -streptavidin tetramer + CD8 + cells, as sampled from blood or with cell-sampling microneedles. (E) OVA-specific IgG titers(log10) as quantified from serum or ISF from sampling microneedles. Data shown are means ± s.e.m. from one representative of two independent experiments. ns, nonsignificant, **, p < 0.01, ****, p < 0.0001, analyzed by one-way ANOVA, followed by Tukey’s HSD.

Journal: Science translational medicine

Article Title: Cell- and Fluid-Sampling Microneedle Patches for Monitoring Tissue-Resident Immunity

doi: 10.1126/scitranslmed.aar2227

Figure Lengend Snippet: (A) Groups of OVA-immunized or naïve C57Bl/6 mice ( n =9/group) were injected intradermally in the ear at time zero with 2 μg OVA and 5 μg each of adjuvants polyI:C and pam3Cys. 60 hrs later, sampling microneedle were applied to the same site for 12 hrs, followed by retrieval and flow cytometry analysis. (B) Sample processing and flow cytometry gating strategy. (C-D) Representative flow cytometry plots ( C ) and quantification from groups of animals ( D ) showing OVA-specific SIINFEKL/H-2K b -streptavidin tetramer + CD8 + cells, as sampled from blood or with cell-sampling microneedles. (E) OVA-specific IgG titers(log10) as quantified from serum or ISF from sampling microneedles. Data shown are means ± s.e.m. from one representative of two independent experiments. ns, nonsignificant, **, p < 0.01, ****, p < 0.0001, analyzed by one-way ANOVA, followed by Tukey’s HSD.

Article Snippet: Flow cytometry: CellTrace CFSE (Thermo C34554), CountBright absolute counting beads (Thermo C36950), CD3 APC (UCHT1, Biolegend), CD4 PE (RPA-T4, Biolegend), CD8 AF700 RPA-T8, Biolegend), CD45 V500 (HI30, BD Horizon), HLA-DR PECy7 (L243, BD Pharmingen), CD11c V450 (B-Ly6, BD Horizon), CD14 PerCPCy5.5 (HCD14, Biolegend).

Techniques: Injection, Sampling, Flow Cytometry

(A) Confocal micrograph of SSMN array surface showing alginate (blue), embedded ICMVs (red), and recruited cells (green), following application to the skin of eGFP mice. Scale bar 20 μm. (B) SEM of SSMNs post-application to mouse skin for 24 hours showing lymphocytes, approximately 5μm in diameter, (C) . (D-G) SSMNs containing 5 μg of polyI:C and DiD-labeled ICMVs encapsulating 2 μg Alexa Fluor555-labeled OVA and 5 μg pam3Cys were applied to the ears of OVA-immunized mice ( n =3/group) for 24 hours, followed by retrieval, antibody staining and phenotypic analysis via imaging cytometry (D). (E) Cell phenotypes obtained from SSMNs. (F) Representative imaging cytometry data showing overlay of fluorescent channels for live/dead dye Sytox, CD45, CD3, CD19, CD11c, DiD (ICMV) and OVA-Alexa Fluor555. Dimension of each well is 50 μm. (G) Representative 2D imaging cytometry plots showing OVA/ICMV fluorescence in CD45 + CD11c + dendritic cells. (H-I) Expression of activation markers CD40 ( H ) and CD86 ( I ) among APCs recovered by microneedles. Data shown are mean ± s.e.m. from one representative of 2-3 independent experiments. *, p<0.05, **, and p<0.01 analyzed by one-way ANOVA, followed by Tukey’s HSD.

Journal: Science translational medicine

Article Title: Cell- and Fluid-Sampling Microneedle Patches for Monitoring Tissue-Resident Immunity

doi: 10.1126/scitranslmed.aar2227

Figure Lengend Snippet: (A) Confocal micrograph of SSMN array surface showing alginate (blue), embedded ICMVs (red), and recruited cells (green), following application to the skin of eGFP mice. Scale bar 20 μm. (B) SEM of SSMNs post-application to mouse skin for 24 hours showing lymphocytes, approximately 5μm in diameter, (C) . (D-G) SSMNs containing 5 μg of polyI:C and DiD-labeled ICMVs encapsulating 2 μg Alexa Fluor555-labeled OVA and 5 μg pam3Cys were applied to the ears of OVA-immunized mice ( n =3/group) for 24 hours, followed by retrieval, antibody staining and phenotypic analysis via imaging cytometry (D). (E) Cell phenotypes obtained from SSMNs. (F) Representative imaging cytometry data showing overlay of fluorescent channels for live/dead dye Sytox, CD45, CD3, CD19, CD11c, DiD (ICMV) and OVA-Alexa Fluor555. Dimension of each well is 50 μm. (G) Representative 2D imaging cytometry plots showing OVA/ICMV fluorescence in CD45 + CD11c + dendritic cells. (H-I) Expression of activation markers CD40 ( H ) and CD86 ( I ) among APCs recovered by microneedles. Data shown are mean ± s.e.m. from one representative of 2-3 independent experiments. *, p<0.05, **, and p<0.01 analyzed by one-way ANOVA, followed by Tukey’s HSD.

Techniques: Labeling, Staining, Imaging, Cytometry, Fluorescence, Expressing, Activation Assay

SSMN arrays containing adjuvants and ICMVs loaded with 2 μg of OVA were applied to the ears of naïve or OVA-immunized C57Bl/6 mice, immunized 8-10 weeks prior, ( n =5/group) for 24 hrs, then retrieved and analyzed via flow cytometry. Shown are experimental timeline (A), representative flow cytometry plots and quantification of OVA-specific CD8 + T cells from blood in naïve (B) and previously immunized (C) mice, before and after SSMN application at day 0 and vaccination on day 24. (D) Serum OVA-specific IgG titers pre- and post-sampling with SSMNs. Data shown are mean ± s.e.m. from one representative of two independent experiments. ****: p < 0.0001 analyzed by one-way ANOVA, followed by Tukey’s HSD.

Journal: Science translational medicine

Article Title: Cell- and Fluid-Sampling Microneedle Patches for Monitoring Tissue-Resident Immunity

doi: 10.1126/scitranslmed.aar2227

Figure Lengend Snippet: SSMN arrays containing adjuvants and ICMVs loaded with 2 μg of OVA were applied to the ears of naïve or OVA-immunized C57Bl/6 mice, immunized 8-10 weeks prior, ( n =5/group) for 24 hrs, then retrieved and analyzed via flow cytometry. Shown are experimental timeline (A), representative flow cytometry plots and quantification of OVA-specific CD8 + T cells from blood in naïve (B) and previously immunized (C) mice, before and after SSMN application at day 0 and vaccination on day 24. (D) Serum OVA-specific IgG titers pre- and post-sampling with SSMNs. Data shown are mean ± s.e.m. from one representative of two independent experiments. ****: p < 0.0001 analyzed by one-way ANOVA, followed by Tukey’s HSD.

Techniques: Flow Cytometry, Sampling

(A-D) Groups of C57Bl/6 mice ( n =5/group) were primed and boosted with OVA and adjuvant, and then repeatedly sampled using SSMN arrays and blood draws over time. (A) Timeline. (B) OVA-specific CD8 + T cells over time. ( C-D ), T RM s ( C ) and OVA-specific T RM s ( D ), quantified from SSMNs and blood. (E-H) Groups of C57Bl/6 mice ( n =5/group) were infected with 2x10 6 PFU of SIVgag-expressing vaccinia virus via tail skin scarification and sampled beginning 11 weeks post infection via blood draws or SSMNs applied to the ear containing ICMVs (2 μg AL11 SIVgag peptide and 5 μg pam3Cys) and 5 μg polyI:C. After 24 hrs, patches were retrieved and analyzed via flow cytometry. Experimental timeline (E) , frequency of SIVgag tetramer + CD8 + cells (F) , enumeration of CD8 + CD69 + CD103 + T RM cells (G) and SIVgag-specific T RM s (H) from blood and SSMNs. (I-K) Timeline (I) , frequency of SIVgag-specific CD8 + T cells (J) and frequency of SIVgag-specific T RM s (K) , when sampled at various times post vaccinia infection. Data shown are mean ± s.e.m. from one representative of 2-3 independent experiments, analyzed by Wilcox-Mann-Whitney test (F-H) , one-way ANOVA (K) or two-way ANOVA (B, J) , ns, nonsignificant, **p < 0.01, ***p < 0.001, ****, p < 0.0001.

Journal: Science translational medicine

Article Title: Cell- and Fluid-Sampling Microneedle Patches for Monitoring Tissue-Resident Immunity

doi: 10.1126/scitranslmed.aar2227

Figure Lengend Snippet: (A-D) Groups of C57Bl/6 mice ( n =5/group) were primed and boosted with OVA and adjuvant, and then repeatedly sampled using SSMN arrays and blood draws over time. (A) Timeline. (B) OVA-specific CD8 + T cells over time. ( C-D ), T RM s ( C ) and OVA-specific T RM s ( D ), quantified from SSMNs and blood. (E-H) Groups of C57Bl/6 mice ( n =5/group) were infected with 2x10 6 PFU of SIVgag-expressing vaccinia virus via tail skin scarification and sampled beginning 11 weeks post infection via blood draws or SSMNs applied to the ear containing ICMVs (2 μg AL11 SIVgag peptide and 5 μg pam3Cys) and 5 μg polyI:C. After 24 hrs, patches were retrieved and analyzed via flow cytometry. Experimental timeline (E) , frequency of SIVgag tetramer + CD8 + cells (F) , enumeration of CD8 + CD69 + CD103 + T RM cells (G) and SIVgag-specific T RM s (H) from blood and SSMNs. (I-K) Timeline (I) , frequency of SIVgag-specific CD8 + T cells (J) and frequency of SIVgag-specific T RM s (K) , when sampled at various times post vaccinia infection. Data shown are mean ± s.e.m. from one representative of 2-3 independent experiments, analyzed by Wilcox-Mann-Whitney test (F-H) , one-way ANOVA (K) or two-way ANOVA (B, J) , ns, nonsignificant, **p < 0.01, ***p < 0.001, ****, p < 0.0001.

Techniques: Adjuvant, Infection, Expressing, Virus, Flow Cytometry, MANN-WHITNEY

Five SSMN arrays each were applied to excised human skin samples from n =5 donors for 16 hrs, and then gel coatings were digested for analysis of recovered cells via flow cytometry. (A) Timeline and photographs of the experimental setup. (B-D) Enumeration of recovered total live cells (B) , CD8 + cells (C) , CD11c + CD45 + cells (D) , per sampling microneedle array, from microneedles containing no adjuvants, or SSMNs containing polyI:C and ICMVs encapsulating pam3Cys. Data shown are mean ± s.e.m. from two independent experiments with samples obtained from 5 total donors, analyzed by two-tailed nonparametric Mann-Whitney test, **p < 0.01.

Journal: Science translational medicine

Article Title: Cell- and Fluid-Sampling Microneedle Patches for Monitoring Tissue-Resident Immunity

doi: 10.1126/scitranslmed.aar2227

Figure Lengend Snippet: Five SSMN arrays each were applied to excised human skin samples from n =5 donors for 16 hrs, and then gel coatings were digested for analysis of recovered cells via flow cytometry. (A) Timeline and photographs of the experimental setup. (B-D) Enumeration of recovered total live cells (B) , CD8 + cells (C) , CD11c + CD45 + cells (D) , per sampling microneedle array, from microneedles containing no adjuvants, or SSMNs containing polyI:C and ICMVs encapsulating pam3Cys. Data shown are mean ± s.e.m. from two independent experiments with samples obtained from 5 total donors, analyzed by two-tailed nonparametric Mann-Whitney test, **p < 0.01.

Techniques: Flow Cytometry, Sampling, Two Tailed Test, MANN-WHITNEY

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celltrace cfse c34554 (Thermo Fisher)

Structured Review

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1) Product Images from "Targeting CD37 promotes macrophage-dependent phagocytosis of multiple cancer cell types and facilitates tumor clearance in mice"

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