Journal: Cancers

Article Title: Characterization of Cellular and Acellular Analytes from Pre-Cystectomy Liquid Biopsies in Patients Newly Diagnosed with Primary Bladder Cancer

doi: 10.3390/cancers14030758

Figure Lengend Snippet: Gallery of representative rare events detected by HDSCA3.0 in PB samples collected from BCa patients prior to cystectomy or ND with no known pathology. ( A – H ) rare cells and ( I ) LEVs. ( A ) DAPI only; ( B ) Vim; ( C ) CD45/CD31; ( D ) Vim|CD45/CD31; ( E ) CK|Vim|CD45/CD31; ( F ) CK|CD45/CD31; ( G ) mes.CTC; ( H ) epi.CTC; ( I ) LEVs (top left: CK only; bottom left: CK|Vim|CD45/CD31; top right: CK|CD45/CD31; bottom right: CK|Vim.)] Blue: DAPI, Red: CK, White: Vim, Green: CD45/CD31. Images taken at 100× magnification. Scale bar = 10 µm.

Article Snippet: Briefly, samples were stained with 2.5 ug/mL of a mouse IgG1 anti-human CD31:Alexa Fluor ® 647 mAb (clone: WM59, MCA1738A647, BioRad, Hercules, CA, USA) and 100 ug/mL of a goat anti-mouse IgG monoclonal Fab fragments (115-007-003, Jackson ImmunoResearch, West Grove, PA, USA), permeabilized using 100% cold methanol, followed by an antibody cocktail consisting of mouse IgG1/Ig2a anti-human cytokeratins (CKs) 1, 4, 5, 6, 8, 10, 13, 18, and 19 (clones: C-11, PCK-26, CY-90, KS-1A3, M20, A53-B/A2, C2562, Sigma, St. Louis, MO, USA), mouse IgG1 anti-human CK 19 (clone: RCK108, GA61561-2, Dako, Carpinteria, CA, USA), mouse anti-human CD45:Alexa Fluor ® 647 (clone: F10-89-4, MCA87A647, AbD Serotec, Raleigh, NC, USA), and rabbit IgG anti-human vimentin (Vim) (clone: D21H3, 9854BC, Cell Signaling, Danvers, MA, USA).

Techniques:

Journal: Cancers

Article Title: Characterization of Cellular and Acellular Analytes from Pre-Cystectomy Liquid Biopsies in Patients Newly Diagnosed with Primary Bladder Cancer

doi: 10.3390/cancers14030758

Figure Lengend Snippet: Morphometric analysis of individual events detected by HDSCA3.0 in PB samples collected from BCa patients prior to cystectomy. ( A ) tSNE plot of rare cellular events depicting the underlying morphological heterogeneity. Each point represents a single cell and is color coded according to its channel-type classification. ( B ) The same tSNE plot color coded according to a distinct cluster number, as determined by a clustering algorithm. The cells group in multiple clusters spanning across classifications. Visualization of the probability density distributions for select morphometric parameters across channel-type classifications: ( C ) cell area, ( D ) cell eccentricity, ( E ) median CK signal intensity, ( F ) median Vim signal intensity, ( G ) median CD45/CD31 signal intensity.

Article Snippet: Briefly, samples were stained with 2.5 ug/mL of a mouse IgG1 anti-human CD31:Alexa Fluor ® 647 mAb (clone: WM59, MCA1738A647, BioRad, Hercules, CA, USA) and 100 ug/mL of a goat anti-mouse IgG monoclonal Fab fragments (115-007-003, Jackson ImmunoResearch, West Grove, PA, USA), permeabilized using 100% cold methanol, followed by an antibody cocktail consisting of mouse IgG1/Ig2a anti-human cytokeratins (CKs) 1, 4, 5, 6, 8, 10, 13, 18, and 19 (clones: C-11, PCK-26, CY-90, KS-1A3, M20, A53-B/A2, C2562, Sigma, St. Louis, MO, USA), mouse IgG1 anti-human CK 19 (clone: RCK108, GA61561-2, Dako, Carpinteria, CA, USA), mouse anti-human CD45:Alexa Fluor ® 647 (clone: F10-89-4, MCA87A647, AbD Serotec, Raleigh, NC, USA), and rabbit IgG anti-human vimentin (Vim) (clone: D21H3, 9854BC, Cell Signaling, Danvers, MA, USA).

Techniques:

Journal: Scientific Reports

Article Title: HLA-DR cancer cells expression correlates with T cell infiltration and is enriched in lung adenocarcinoma with indolent behavior

doi: 10.1038/s41598-021-93807-3

Figure Lengend Snippet: Mass cytometry antibody panel for lung adenocarcinoma.

Article Snippet: CD45 , 154-Sm , Surface , HI30 , Fluidigm , 3154001B.

Techniques: Mass Cytometry

Journal: Cells

Article Title: SOCS3 Regulates Dectin-2-Induced Inflammation in PBMCs of Diabetic Patients

doi: 10.3390/cells11172670

Figure Lengend Snippet: Antibodies list.

Article Snippet: Anti-CD45 VioBlue , IgG2a , 130-113-684 , Miltenyi Biotec.

Techniques:

Journal: Cells

Article Title: SOCS3 Regulates Dectin-2-Induced Inflammation in PBMCs of Diabetic Patients

doi: 10.3390/cells11172670

Figure Lengend Snippet: Increased Dectin-2 expression in diabetic patients’ PBMCs. Flow cytometric analysis was conducted to identify the expression levels of Dectin-2 on different leukocyte subsets in PBMCs isolated from non-diabetic and diabetic patients. ( A ) Representative dot plot identifying three leukocyte subsets. ( B ) Representative dot plot of Dectin-2 expression in different leukocyte subsets. ( C ) Bar graph analysis of Dectin-2 expression calculated as MFI with representative histogram. ( D ) Representative density plot analysis of CD45 + CD14 + Dectin-2 + population in PBMCs isolated from non-diabetic and diabetic patients. ( E ) The percentage of CD45 + CD14 + Dectin-2 + population in PBMCs isolated from non-diabetic and diabetic patients. ( F ) Bar graph analysis of CD45 + CD14 + Dectin-2 + expression calculated as MFI with representative histogram. Gray (non-stained), green (lymphocytes), blue (monocytes), and purple (neutrophils). Data are presented as mean ± SEM with a minimum of n = 3. Statistical significance was assessed by Student’s t -test. * p < 0.05, ** p < 0.01, and *** p < 0.001.

Article Snippet: Anti-CD45 VioBlue , IgG2a , 130-113-684 , Miltenyi Biotec.

Techniques: Expressing, Isolation, Staining

Journal: The Journal of Clinical Investigation

Article Title: S1PR4 ablation reduces tumor growth and improves chemotherapy via CD8 + T cell expansion

doi: 10.1172/JCI136928

Figure Lengend Snippet: (A) Experimental outline of the AOM/DSS model applied to WT and S1PR4-KO mice. (B) Weight of AOM/DSS-treated WT and S1PR4-KO mice as a percentage of weight at the initiation of treatment (n = 9). (C and D) Relative amounts of CD45+ leukocytes (C) and immune cell populations (D) in the LP of WT and S1PR4-KO mice at day 0 (n = 6), day 8 (n = 8), day 15 (n = 4), and day 84 (n = 9) analyzed by flow cytometry. (E) Representative pictures of WT and S1PR4-KO AOM/DSS-treated colon tissue at day 8 stained with H&E. Scale bars: 1 mm; scale bars of magnified areas:100 μm. (F) Colon weight-to-length ratio determined for WT and KO AOM/DSS-treated mice at days 0 (WT: n = 7, KO: n = 8), 8 (n = 4), 15 (n = 5), and 84 (n = 10). Means ± SEM; 2-tailed Student’s t test; *P < 0.05.

Article Snippet: Primary PyMT tumor sections were stained without antigen retrieval with CD8 antibody (53-6.7, BD Biosciences) or anti-CD45 (D3F8Q, Cell Signaling Technology) overnight at 4°C and with corresponding secondary anti–rat IgG-HRP (sc-2006, Santa Cruz Biotechnology) for 1 hour at 4°C.

Techniques: Flow Cytometry, Staining