Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Contrived samples in HDB. (A) Representative on-chip immunofluorescence image of cells captured with the cartridge of the Hitachi MCA system. Red: CD45; blue: DAPI; green: cytokeratin, white: brightfield image of 8 µm × 100 µm microcavity, imaged at 100×. CTCs are defined as cells that lack CD45 but have a defined DAPI nuclear stain and cytokeratin. (B) Recovery of tumor cell lines spiked into healthy donor blood. The recovery rates of spiked cell lines were 91.5% for H358 and 84.3% for A549. HDB, healthy donor blood; MCA, Micro Cavity Array.

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques: Immunofluorescence, Staining

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Image of CD45 depletion. (A) Shows an MCA cartridge imaged by immunofluorescence where the leukocytes were depleted using CD45 Microbeads prior to enrichment. (B) Shows high levels of co-eluting leukocytes in an MCA cartridge without pre-treatment with CD45 microbeads. MCA, Micro Cavity Array.

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques: Immunofluorescence

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Multivariate analyses

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques:

Journal: Translational Lung Cancer Research

Article Title: Enumeration and molecular characterization of circulating tumor cells enriched by microcavity array from stage III non-small cell lung cancer patients

doi: 10.21037/tlcr-20-841

Figure Lengend Snippet: Multivariate analyses with CTC count

Article Snippet: To generate this leukocyte-poor fraction, we incubated peripheral blood samples with StraightFrom Whole Blood CD45 MicroBeads at 50 μL per mL of blood passed through a magnetic column (Miltenyi Biotec) before enrichment by the MCA system.

Techniques:

Journal: Nature medicine

Article Title: Cranioencephalic functional lymphoid units in glioblastoma.

doi: 10.1038/s41591-024-03152-x

Figure Lengend Snippet: Fig. 2 | CB cellular immune profile. a, Schematic depicts sources of CD45+ immune cells. b, UMAP of integrated scRNA-seq data from CD45 (PTPRC)- expressing immune cells. Tissue sources color coded, numbers (n) of biosamples indicated per source. Insets visualize expression of selected genes. c, Overlay of SingleR- and marker-based annotation of cell types. d, Bubble plot summarizing prevalence of immune cell subsets among CD45+ nongranulocytes, by flow cytometry. e, Scatter plot of scVDJ data from n = 3 patients with glioblastoma visualizing shared T cell clonotypes between CB and tumor. Clone size visualized by number of cells per clone, each point represents a unique clone. Axes, log- transformed counts of cells (log1p). Exclusive CB or tumor clones plotted along the y and x axis, respectively. Shared clones located in the central area of the graph. f, Top ten differentially expressed genes (ranked by log2(FC)) comparing tumor-shared expanding clonotypes versus nonexpanding singlets in the CB. DEGs detected by FindMarkers() Seurat function, per default setting (two-sided

Article Snippet: Selection and preservation of immune cells Single-cell suspensions from tumor tissue, bone and blood were enriched for vital CD45+ cells by the REAlease CD45 (TIL) MicroBead Kit (following the manufacturer’s protocol; Miltenyi Biotec, 130-121- 563).

Techniques: Expressing, Marker, Flow Cytometry, Transformation Assay, Clone Assay

Journal: Nature Communications

Article Title: Alternations in inflammatory macrophage niche drive phenotypic and functional plasticity of Kupffer cells

doi: 10.1038/s41467-024-53659-7

Figure Lengend Snippet: a, b Schematic of the experimental liver metastasis (LvMet) models used and flow cytometry (FC) quantification of CD45 + leukocytes in LvMet and adjacent normal (adj. Normal) tissues from the same livers ( n = 5 for MC38; n = 5 for E0771; n = 6 for hepatocellular carcinoma, HCC). c Multi-colour FC analysis of immune cells in LvMet and adj. Normal tissues from the same livers ( n = 5 for MC38; n = 5 for E0771; n = 4 for HCC). d Schematic diagram and experimental workflow for CITE-seq. e . UMAP embedding of N-macs and LMAMs, and feature plots showing the expression of KC-identity genes and mo-mac signature genes. f Schematic design of dual-fluorescent reporter mice for tracing macrophages. g–i Representative immunofluorescence (IF) staining and FC quantification of KC markers Clec4f and Timd4 in adj. Normal and LvMet tissues from the same livers ( n = 6 for E0771; scale bar, 50 μm). j Representative IF staining and quantification of Timd4 expression in liver metastasis tissue samples derived from breast cancer patients (triple negative, n = 3; HER2 + , n = 3; scale bar, 50 μm). Mean ± s.e.m. shown. P values were calculated by comparing individual animals using two-tailed paired ( b, g– j ) Mann–Whitney U -test.

Article Snippet: The CD45 Microbeads (RWD Life Science, #K1304) were used to enriched immune cells from single cell suspension through magnetic cell separation.

Techniques: Flow Cytometry, Expressing, Immunofluorescence, Staining, Derivative Assay, Two Tailed Test, MANN-WHITNEY

Journal: Nature Communications

Article Title: Alternations in inflammatory macrophage niche drive phenotypic and functional plasticity of Kupffer cells

doi: 10.1038/s41467-024-53659-7

Figure Lengend Snippet: a Schematic design of KC lineage tracing mice. b IF quantification of KC-derived cells (HA tag + or sfGFP + ) in adjacent normal tissues and LvMet tissues from the same livers ( n = 5 for MC38; n = 5 for E0771; scale bar, 50 μm). c FC analysis of sfGFP and tdT expression in HelixNIR- CD45 + F4/80+ cells ( n = 5 for MC38; n = 5 for E0771). d,e Schematic and experimental design of monocyte lineage tracing and genetic ablation (IF, immunofluorescence; bw, body weight; og, oral gavage). f-h . Representative histogram and IF image, as well as FC quantification of tdT expression in N-macs and LMAMs from the same livers (n = 7 for MC38; n = 5 for E0771; scale bar, 50 μm). i IF staining of LMAMs upon acute monocyte depletion ( Cx3cr1 CreERT2 ;R26 LSL-DTA , n = 5 for MC38, n = 6 for E0771; littermate control Cx3cr1 CreERT2 ;R26 WT , n = 4 for MC38; n = 6 for E0771; scale bar, 50 μm). j FC quantification of LMAMs upon acute monocyte depletion ( Cx3cr1 CreERT2 ;R26 LSL-DTA , n = 5 for MC38, n = 6 for E0771; littermate control Cx3cr1 CreERT2 ;R26 WT , n = 5 for MC38; n = 5 for E0771). k IF quantification of monocytes and LMAMs by impairing monocyte chemotaxis ( Ccr2 GFP/GFP : n = 6 for MC38; n = 5 for E0771; Ccr2 GFP/WT littermates: n = 5 for MC38; n = 5 for E0771; scale bar, 50 μm). l Representative FC plot showing loss of Ccr2 expression in LMAMs of Ccr2 GFP/GFP mice, and quantification of the percents of monocyte and LMAM in CD45 + leukocytes ( Ccr2 GFP/GFP : n = 6 for MC38; n = 8 for E0771; Ccr2 GFP/WT littermates: n = 4 for MC38; n = 8 for E0771). Mean ± s.e.m. shown. Smaller dots are values from individual fields. Outlined circles are mean values taken over multiple fields/sections from the same mouse. P values were calculated by comparing individual animals using two-tailed paired ( b, c, h ) and unpaired ( i - l ) Mann–Whitney U -test.

Article Snippet: The CD45 Microbeads (RWD Life Science, #K1304) were used to enriched immune cells from single cell suspension through magnetic cell separation.

Techniques: Derivative Assay, Expressing, Immunofluorescence, Staining, Control, Chemotaxis Assay, Two Tailed Test, MANN-WHITNEY