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3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 <t>(purple),</t> <t>Collagen</t> <t>III</t> (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.
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3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 <t>(purple),</t> <t>Collagen</t> <t>III</t> (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.
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Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and <t>Collagen</t> <t>III</t> quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.
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Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and <t>Collagen</t> <t>III</t> quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.
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Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and <t>Collagen</t> <t>III</t> quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.
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Image Search Results


3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 (purple), Collagen III (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.

Journal: STAR Protocols

Article Title: Protocol for 3D-guided sectioning and deep cell phenotyping via light sheet imaging and 2D spatial multiplexing

doi: 10.1016/j.xpro.2025.104296

Figure Lengend Snippet: 3D light sheet and 2D multi-cyclic imaging data comparison (Human OvCa) (A) Imaris 3D surface rendering of autofluorescence (cyan) and CD326 positive cells (red). (B) Imaris 3D surface rendering of autofluorescence (cyan) with target plane in yellow. (C) Light sheet guided target plane selection representing CD326 positive cell (purple), CD45 positive cells (red), and CD3 positive cells (green). (D) DAPI overview image of selected tissue slice for 2D MACSima™ imaging. (E) Magnified merged six color multiparameter MICS image with CD45 (green), CD326 (cyan), FOLR1 (purple), Collagen III (red), Collagen IV (red), and CD31 (yellow). (F–L) Single staining MICS images (white) of DAPI (F), CD45 (G), CD326 (H), FOLR1 (I), Collagen III (J), Collagen IV (K), and CD31 (L) (gray) (see “Antibodies”). Scale bars: (A–F) 1 mm; (E) 250 μm; (F–L) 500 μm.

Article Snippet: Collagen III antibody, anti-human, PE, REAdye_lease , Miltenyi Biotec B.V. & Co. KG , Cat# 130-127-357 RRID: AB_2905171.

Techniques: Imaging, Comparison, Selection, Staining

Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and Collagen III quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Journal: bioRxiv

Article Title: TLR5 drives metabolic dysfunction-associated steatohepatitis through lipid- and flagellin-induced hepatocyte injury signalling

doi: 10.64898/2026.02.05.703969

Figure Lengend Snippet: Schematic representation of precision-cut liver slices (PCLS) setup, created in Biorender. PCLS were cultured for 16 hours in the presence or absence of 1µM TH1020 (TLR5 inhibitor) and treated with bovine serum albumin (BSA) or oleic acid and palmitic acid (OA PA) for 24 hours (n=3). Representative immunohistochemistry images H&E and Collagen III quantification (% area) of PCLS shown. Scale bar 100µM and 50µM. Error bars represent mean ± standard deviation; P values determined by t test or ANOVA with post-test multiple comparisons. ANOVA, analysis of variance. Only statistically significant comparisons (p<0.05) are highlighted.

Article Snippet: FFPE tissue sections were immunostained with rabbit polyclonal antibody directed against human Collagen Type III (rabbit IgG, ref: 22734-1-A, Proteintech, 1/2000) using the fully automated BOND Max system from Leica Microsystems and a BOND Polymer Refine Detection kit (#DS9800).

Techniques: Cell Culture, Immunohistochemistry, Standard Deviation