Journal: Cancer Science

Article Title: In vivo subcellular imaging of tumors in mouse models using a fluorophore-conjugated anti-carcinoembryonic antigen antibody in two-photon excitation microscopy

doi: 10.1111/cas.12500

Figure Lengend Snippet: Inoculation of human cancer cells into immunodeficient mice and in vivo macroscopic imaging using a fluorescence zoom microscope. (a) Schema of the sites of inoculation of human cancer cells. The cells were inoculated s.c. into the back skin of nude mice at the rostral–ventral site (HT1080-GFP cells), the caudal–ventral site (HT1080-GFP-CEA cells) or the dorsal site (MKN45-GFP cells). (b) Schema of preparation of skin flaps. Seven or eight days after the inoculation, the inoculation sites were exposed by the skin-flap method. (c–e) In vivo macro imaging of tumors. In vivo macro imaging of the tumor masses was performed using a fluorescence zoom microscope 24 h after injection of Alexa Fluor 594-conjugated anti-CEA antibody (50 μg/mouse). Exposure times for the GFP and Alexa Fluor 594 fluorescence images were 30 and 100 ms, respectively. These experiments were repeated three times and similar results were obtained.

Article Snippet: In vivo macroscopic imaging was performed using a fluorescence zoom microscope (MULTIZOOM AZ100; Nikon) with a GFP-HQ filter set (for GFP) and a Texas Red filter set (for Alexa Fluor 594), and an AZ-Plan Apo 0.5× Plan Fluor objective lens (Nikon).

Techniques: In Vivo, Imaging, Fluorescence, Microscopy, Injection

Journal: Cancer Science

Article Title: In vivo subcellular imaging of tumors in mouse models using a fluorophore-conjugated anti-carcinoembryonic antigen antibody in two-photon excitation microscopy

doi: 10.1111/cas.12500

Figure Lengend Snippet: In vivo fluorescence imaging using a two-photon microscope. After the in vivo macroscopic imaging as shown in Figure (c–e), the same tumors were observed by two-photon excitation microscopy. (a–f) 3-D and 2-D images of HT1080-GFP (a, d), HT1080-GFP-CEA (b, e) and MKN45 (c, f) cells were acquired by two-photon excitation microscopy. Each 2-D image represents an orthogonal view: x-y (center panel), y-z (left panel) and x-z (lower panel). Red, green and blue indicate Alexa Fluor 594 fluorescence, GFP fluorescence and second harmonic generation (SHG), respectively. (g–i) Magnified images of (d–f).

Article Snippet: In vivo macroscopic imaging was performed using a fluorescence zoom microscope (MULTIZOOM AZ100; Nikon) with a GFP-HQ filter set (for GFP) and a Texas Red filter set (for Alexa Fluor 594), and an AZ-Plan Apo 0.5× Plan Fluor objective lens (Nikon).

Techniques: In Vivo, Fluorescence, Imaging, Microscopy

Journal: Cancer Science

Article Title: In vivo subcellular imaging of tumors in mouse models using a fluorophore-conjugated anti-carcinoembryonic antigen antibody in two-photon excitation microscopy

doi: 10.1111/cas.12500

Figure Lengend Snippet: In vivo fluorescence macroscopic and microscopic imaging of lymph-node metastases by a fluorescence zoom microscope and a two-photon excitation microscope. (a–c) A footpad spontaneous metastasis model using HT1080-GFP-CEA cells observed by a fluorescence zoom microscope. The popliteal lymph node was exposed, and multiple images were collected: bright field image (a), GFP (b) and Alexa Fluor 594 (c). Exposure times for the GFP and Alexa Fluor 594 images were 1000 and 3000 ms, respectively. (d–f) Two-photon excitation microscopy of the popliteal lymph node. After in vivo macroscopic imaging, the same lymph node was observed using a two-photon excitation microscope. Acquired images are shown as 3-D construction (d), cropped 3-D image of (e) and magnified image of (f), respectively. Red, green and blue indicate Alexa Fluor 594 fluorescence, GFP fluorescence and second harmonic generation (SHG), respectively.

Article Snippet: In vivo macroscopic imaging was performed using a fluorescence zoom microscope (MULTIZOOM AZ100; Nikon) with a GFP-HQ filter set (for GFP) and a Texas Red filter set (for Alexa Fluor 594), and an AZ-Plan Apo 0.5× Plan Fluor objective lens (Nikon).

Techniques: In Vivo, Fluorescence, Imaging, Microscopy

Journal: Cell Reports

Article Title: R-spondin1 Controls Muscle Cell Fusion through Dual Regulation of Antagonistic Wnt Signaling Pathways

doi: 10.1016/j.celrep.2017.02.036

Figure Lengend Snippet: Rspo1-null Muscles Show Enhanced Regeneration Caused by a Delay of Muscle Progenitor Cell Differentiation and an Improved Fusion (A) LAMININ (red) and MYOGENIN (green) immunolocalization in TA muscles at 4 d.p.i. (B–D) Quantification of the number of PAX7-positive cells (B), MYOGENIN-positive cells (C), and myonuclei (D) per myofiber 4 d.p.i. (E) DYSTROPHIN (red) and MYOGENIN (green) immunolocalization at 7 d.p.i. (F) Mean CSA of myofibers in TA muscles at 7 d.p.i. (G) Quantification of the number of MYOGENIN-positive cells inside the myofibers showing a higher proportion of fused nuclei at 7 d.p.i. in Rspo1-null mice. (H) Quantification of the number of PAX7-positive cells per myofiber at 7 d.p.i. (I and J) LAMININ (red) immunolocalization on muscle sections at 62 d.p.i. (I) and a whole cross-section of regenerated 62 d.p.i. muscles (J). Images are virtual slides automatically assembled by the Axio Scan.Z1 microscope. (K and L) Quantification of TA muscle weights (K) and mean CSA (L) at 62 d.p.i. (M and N) Number of myofibers per surface unit (N) and of nuclei per myofiber (M) at 62 d.p.i. (O) Distribution of the percentage of myofibers depending on their nuclear number at 62 d.p.i. (P and Q) Fatigue resistance index (P) and specific maximal force (Q) of 62 d.p.i. muscles. Nuclei are stained with Hoechst (blue). Scale bars, 50 μm (A and E); 35 μm (I); 150 μm (J). Error bars indicate SD. ∗ p value < 0.05; ∗∗ p value < 0.01.

Article Snippet: Immunofluorescent stainings were analyzed with an Olympus BX63F microscope, Zeiss Axio Observer.Z1 microscope and AZ100 Nikon Macroscope from Cochin Institute, and Zeiss Axiovert 200M microscope and Axio Scan.Z1 microscope from the ICM.

Techniques: Cell Differentiation, Microscopy, Staining