Journal: PLoS ONE

Article Title: High-Sensitivity Real-Time Imaging of Dual Protein-Protein Interactions in Living Subjects Using Multicolor Luciferases

doi: 10.1371/journal.pone.0005868

Figure Lengend Snippet: (A) Characterization of the probes. a and b: COS-7 cells were transfected with the plasmids, Smad4-McLuc1 plus FLucN-Smad1 or Smad4-McLuc1 plus CBRN-Smad1, in the absence (Mock) and presence of either ALK3CA or ALK3DN receptor. Smad1(AXA) and Smad2(AXA) indicate double mutants of Smad1(S462A,S464A) and Smad2(S465A,S467A), respectively. The cells were incubated for 12–16 h and the luciferase activities were measured. c: COS-7 cells were transfected with the plasmids, ELucN-Smad2 plus Smad4-McLuc1 or ELucN-Smad2(AXA) plus Smad4-McLuc1 in the absence (Mock) and presence of either ALK5CA or ALK5DN receptor. d: COS-7 cells transfected with the plasmids FLucN-Smad1 and Smad4-McLuc1 were stimulated with BMP-2 (50 nM) for 2 h and luciferase activities were measured. Error bars represent s.d. calculated for three independent samples. e: Western blotting analysis of the expression of Smad1–Smad4 or Smad2–Smad4 protein in the presence of ALK3CA, ALK3DN, ALK5CA or ALK5DN. (*P<0.05, **P<0.01, ***P<0.001) (B)–(D) Real-time bioluminescence images of Smad1–Smad4 and Smad2–Smad4 interactions using CBRN-Smad1 and Smad4-McLuc1 (B), CBRN-Smad2 and Smad4-McLuc1 (C), and CBRN-Smad1, ELucN-Smad2 and Smad4-McLuc1 (D), in a Xenopus embryo. RNAs encoding the Smad probes were injected into the animal pole of a 2-cell stage Xenopus embryo. The embryo was incubated for 24 h at 13°C and thereafter, digital images of Venus (gray) and bioluminescence (pseudocolor) were acquired using a microscopic system equipped with an EM-CCD camera. Bar, 1 mm.

Article Snippet: Ten minutes after injection of D-luciferin, bioluminescence images were taken using a cooled CCD camera (Versarray 1300B, Princeton Instruments Inc.) with or without a filter (BP(ELuc); 520±25 nm, BP(SLRLuc); 630±37.5 nm, Chroma Technology Corp.).

Techniques: Transfection, Incubation, Luciferase, Western Blot, Expressing, Injection

Journal: PLoS ONE

Article Title: High-Sensitivity Real-Time Imaging of Dual Protein-Protein Interactions in Living Subjects Using Multicolor Luciferases

doi: 10.1371/journal.pone.0005868

Figure Lengend Snippet: (A) Results of analyses of interactions between 14-3-3 and Bad mutants. The COS-7 cells were transiently transfected with Bad–McLuc1 and ELucN–14-3-3; luciferase activities were tested. Error bars represent s.d. calculated for three independent samples. (B) Results of Western blotting analysis of Bad phosphorylation. Expression levels of Bad and its mutants were determined using immunoblotting with the anti-V5 antibody (left). Mutations of Bad at S112A, S136A, and S155A were confirmed by immunoblotting with the respective antibodies (right). (C) An inhibitory effect of antimycin or HA14-1 on the bioluminescence was developed by the Bad-Bcl-2 interaction. Antimycin (10 µM) or HA14-1 (10 µM) was added to the cells after transfection and incubated for 20 h. The cells were harvested and the photon counts were analyzed. Error bars represent s.d. calculated for three independent samples. (D) Dual color imaging of mice with a single substrate. The images shown are superimposed on the optical CCD bioluminescence image without a filter (Open) or with a band-pass filter of BP(ELuc) (525±25 nm) or BP(SLRLuc) (630±37.5 nm). A nude mouse was imaged after implantation of COS-7 cells that had been transiently transfected with plasmids SLRLuc (site 1), Bad–McLuc1 and ELucN–14-3-3 (site 2), SLRLuc plus Bad–McLuc1 and ELucN–14-3-3 (site 3), and SLRLuc plus Bad(S112A, A136A, A155A) –McLuc1 and ELucN–14-3-3 (site 4). To obtain photon flux information from mice, the bioluminescence intensity was shown as pseudocolors. Photon counts with BP(ELuc) divided by those with BP(SLRLuc) are shown at the right side of the image.

Article Snippet: Ten minutes after injection of D-luciferin, bioluminescence images were taken using a cooled CCD camera (Versarray 1300B, Princeton Instruments Inc.) with or without a filter (BP(ELuc); 520±25 nm, BP(SLRLuc); 630±37.5 nm, Chroma Technology Corp.).

Techniques: Transfection, Luciferase, Western Blot, Phospho-proteomics, Expressing, Incubation, Imaging

Journal: Antioxidants

Article Title: Imaging of Lipid Peroxidation-Associated Chemiluminescence in Plants: Spectral Features, Regulation and Origin of the Signal in Leaves and Roots

doi: 10.3390/antiox11071333

Figure Lengend Snippet: Stability of lipid peroxidation-related SCL. ( a ) Time course of the change in endogenous chemiluminescence in wounded Arabidopsis leaves: in red, total emission; in blue, emission at wavelengths <600 nm. ( b ) Comparison of the time dependence of SCL from wounded WT and scl14 leaves. ( c ) Time course of the changes in luminescence emission from oxidized lipids in the blue-green (<600 nm) and red/far-red (>640 nm) light domains. Linolenic acid was oxidized by soybean LOX as in e–g. The blue-green and (far-)red signals were measured simultaneously by placing a LG640 filter or a LS600 filter above the lipid solutions in the black box of the CCD camera installation.

Article Snippet: The luminescence emissions from plants, leaf discs or liquid solutions were imaged using a highly sensitive, liquid N 2 -cooled charge coupled device (CCD) camera (VersArray 1300 B, Roper Scientific).

Techniques: Comparison

Journal: International Journal of Biomedical Imaging

Article Title: Cerenkov Luminescence Tomography for In Vivo Radiopharmaceutical Imaging

doi: 10.1155/2011/641618

Figure Lengend Snippet: Schematic diagram of our imaging system. In vivo imaging system consists of a CCD camera, small animal anesthesia machine, a Micro-focus X-ray source, an X-ray flat panel detector, a rotation stage, and an antiradiation shield.

Article Snippet: The CCD camera (Princeton Instruments VersArray 1300 B, Roper Scientific, Trenton, NJ) has 1340 × 1300 pixels with 20 × 20 μ m-sized pixels.

Techniques: Imaging, In Vivo Imaging

Journal: International Journal of Biomedical Imaging

Article Title: Cerenkov Luminescence Tomography for In Vivo Radiopharmaceutical Imaging

doi: 10.1155/2011/641618

Figure Lengend Snippet: System block diagram of the in vivo imaging system, developed by ourselves. The region near the bladder belongs to the view of the CCD camera.

Article Snippet: The CCD camera (Princeton Instruments VersArray 1300 B, Roper Scientific, Trenton, NJ) has 1340 × 1300 pixels with 20 × 20 μ m-sized pixels.

Techniques: Blocking Assay, In Vivo Imaging

Journal: International Journal of Biomedical Imaging

Article Title: Cerenkov Luminescence Tomography for In Vivo Radiopharmaceutical Imaging

doi: 10.1155/2011/641618

Figure Lengend Snippet: The views of the mouse bed with the fixed markers. The healthy mouse was injected with Fenestra LC and FDG. The rotation stage is set to rotate at 0°, 90 °, 180°, and 270° for taking photos respectively. The CCD camera was fixed in front of the mouse bed.

Article Snippet: The CCD camera (Princeton Instruments VersArray 1300 B, Roper Scientific, Trenton, NJ) has 1340 × 1300 pixels with 20 × 20 μ m-sized pixels.

Techniques: Injection