ivis kinetic imaging system  (Caliper Life Sciences)

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Schematic showing comparison of AAV9 vectors packaging either the strong, constitutively active chicken beta actin (CBA) promoter or minimal Heat shock protein 1a ( Hsp1a ) promoter to direct fLuc expression. n=5 mice/AAV group. ( B ) Representative IVIS images of mice injected with AAV containing either CBA (top) or Hsp1a (bottom) promoters. Red boxes indicate regions of interest (ROIs) marking cardiac, liver, and whole-body expression. ( C ) Average radiance measured from cardiac ROIs from CBA (top) or Hsp1a (bottom) promoters shows relatively consistent expression from 30 to 68 days post-AAV injection. Square, male mice. Circle, female mice. ( D, E ) Average radiance measured from liver ( D ) and whole-body ( E ) ROIs showed relatively consistent levels of expression over time for both promoters.

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: Comparison, Expressing, Injection

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Schematic illustration of study design. Albino BALB/c mice were systemically injected with AAV9 vectors packaging fLuc reporter cassettes directed by TREEs and a permissive promoter. Mice underwent ischemia/reperfusion (I/R) surgery at D61 and were imaged by IVIS at indicated time points. ( B ) Representative IVIS images indicate changes of expression over time and space for each vector. Cardiac region of interest (ROI) indicated by red box. n=2 mice. ( C ) Average radiance measured from cardiac ROIs plotted over days post-injury (dpi). Average radiance normalized to their baseline pre-injury was also plotted (right). n=2 mice. ( D ) Average cardiac radiance showed a transient increase in expression for both REN (left) and 2ankrd1aEN (right) after I/R injury, whereas sham-operated animals showed relatively constant expression. n=2 mice for I/R, n=3 mice for sham.

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: Injection, Expressing, Plasmid Preparation

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Schematic of experimental timeline, comparing AAV9 and AAV.cc84 capsids for systemic delivery of REN-Hsp1a:: fLuc. Mice were in vivo bioluminescence imaging (IVIS) imaged in the weeks following AAV delivery and post-sham surgery. ( B ) Representative IVIS images of mice injected with either AAV9 (left) or AAV.cc84 (right) at 14 (top) and 21 days (bottom) post-AAV injection. Mice were also subdivided by biological sex to account for sex differences in AAV liver tropism. ( C ) Average radiance from liver regions of interest (ROIs) showed significantly higher expression in AAV9-transduced mice compared to AAV.cc84 through all timepoints (n=6 mice, Holm–Sidak multiple comparisons test). ( D ) Representative IVIS images of harvested organs at 42 days post-AAV demonstrate liver expression with AAV9 (left) while undetected with AAV.cc84 (right). ( E ) Vector genome quantification from collected liver samples reveals higher liver transduction with AAV9 compared to AAV.cc84 for both female and male mice.

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: In Vivo, Imaging, Injection, Expressing, Plasmid Preparation, Transduction

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Representative in vivo bioluminescence imaging (IVIS) images of mice injected with either AAV9 (left) or AAV.cc84 (right) at 7 (top) and 35 days (bottom) post-AAV injection. ( B ) Average radiance in the heart was similar between AAV9 and AAV.cc84 (n=6 mice, Holm–Sidak multiple comparisons test). ( C ) Vector genome quantification from cardiac tissues showed similar levels of vector genomes between AAV9 and AAV.cc84 for both sexes (n=3 mice, Holm–Sidak multiple comparisons test). ( D ) Representative IVIS images of mice with regions of interest (ROIs) used to measure expression in head/neck and lower abdomen (white dashed box). ( E ) Average radiance measured in the head/neck (left) and lower abdomen (right) was similar between AAV9 and AAV.cc84 over the course of the study (n=6 mice, Holm–Sidak multiple comparisons test).

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: In Vivo, Imaging, Injection, Plasmid Preparation, Expressing

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Schematic of experimental timeline comparing expression between Hsp1a and 2ankrd1aEN when delivered at 4 dpi. ( B ) Representative in vivo bioluminescence imaging (IVIS) images of mice injected with Hsp1a (top) or 2ankrd1aEN- Hsp1a (bottom) after ischemia/reperfusion (I/R) injury. ( C ) Cardiac average radiance normalized to the 7 dpi time point increased over time with 2ankrd1aEN while remaining stable with Hsp1a (n=4 mice, Holm–Sidak multiple comparisons test). ( D ) Average cardiac radiance directed by 2ankrd1aEN was significantly higher in mice with I/R injury compared to sham at 7 dpi (n=4 mice, Welch’s t -test). ( E ) Average cardiac radiance was more significantly elevated in the first 21 days post-injury in mice with I/R injury compared to sham (n=4 mice, Mann–Whitney tests).

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: Expressing, In Vivo, Imaging, Injection, MANN-WHITNEY

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Ischemia/reperfusion (I/R) surgery injury extent was assessed by ejection fraction via echocardiography to estimate injury prior to AAV delivery (n=4 mice). ( B ) Cardiac average radiance plotted for each individual mouse with I/R injury plotted over time with either Hsp1a:: fLuc (gray) or 2ankrd1aEN- Hsp1a:: fLuc (black). ( C ) Representative in vivo bioluminescence imaging (IVIS) images of sham-operated mice injected with AAV.cc84 packaged with 2ankrd1aEN- Hsp1a:: fLuc.

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: In Vivo, Imaging, Injection

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Representative in vivo bioluminescence imaging (IVIS) images of mice with sham (top) or (bottom) surgery transduced with either AAV9 (left) or IR41 (right) packaged with 2ankrd1aEN- Hsp1a:: fLuc (n=3–4 mice). ( B, C ) Cardiac average ( B ) and maximum ( C ) radiance was elevated in MI mice transduced with IR41 compared to AAV9 (n=3–4 mice, Holm–Sidak multiple comparisons test). ( D ) Viral genome quantification from heart tissues was elevated in MI mice injected with IR41 compared to AAV9 (n=3–4 mice, Holm–Sidak multiple comparisons test).

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: In Vivo, Imaging, Transduction, Injection

Journal: eLife

Article Title: Spatial and longitudinal tracking of enhancer-AAV vectors that target transgene expression to injured mouse myocardium

doi: 10.7554/eLife.107148

Figure Lengend Snippet: ( A ) Compiled in vivo bioluminescence imaging (IVIS) images of mice that underwent sham or MI surgery with AAV9 or IR41 transduced at 3 dpi. Mice were imaged at 7, 14, and 21 dpi. ( B, C ) Cardiac average ( B ) and maximum ( C ) radiance were statistically similar between AAV9 and IR41 in sham-operated mice.

Article Snippet: Bioluminescence images were then collected using an IVIS Kinetic instrument and Living Image Software (PerkinElmer).

Techniques: In Vivo, Imaging