Imaging of Vascular Anatomy in C57Bl/6 MiceThis	  case	  study	  provides	  guidance	  and	  recommendations	  for	  conducting	  vascular	  imaging	  studies	  in	  normal	  mice	  with	  Fenestra	  VC.	  You	  should	
  note	  that	  other	  examples	  of	  the	
  capabilities	  of	  Fenestra	  and	  a	  selection	  of	  additional	  case	  studies	  are	  available	  on	  the	  MediLumine	  website	  at	  http://www.medilumine.com/resources	  	  	  Fenestra®	  VC	  provides	  flexible,	  long-­‐lasting	  contrast	
  enhancement	  for	  a	  wide	
  range	  of	  computed	  tomography	  imaging	  applications,	  including	  vascular	  and	  hepatobiliary	  anatomy	  and	  function.	  	  Animal	  Model	  StrainC57BI/6	  (31	  to	  36	  g	  males).	  

Model
Normal	  mice	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  Animal	
  Preparation	  AnesthesiaMice	  were	  anesthetized	  with	  an	  intraperitoneal	  injection	  of	  a	  mixture	  of	  ketamine	
  (80	  mg/kg	  body	  weight)	  and	  xylazine	  (5	  mg/kg	  body	  weight),	  which	  afforded	  45	  to	  60	  minutes	  of	  anesthesia.	  Anesthesia	  was	  maintained	  with	  quarter	  dose	  increments	  during	  the	  duration	  of	  the	  study.	  

AdministrationFenestra®	  VC	  was	  injected	  intravenously	
  into	  the	  lateral	
  tail	  vein	  of	  anesthetized	  mice	  at	  a	  dose	  of	  0.4	  ml	  per	  20	  g	  body	  weight	  over	
  a	
  period	  of	  30	  to	  60	  seconds.	
  A	  1	  ml	  disposable	  syringe	  fitted	  with	  a	  30-­‐gauge	  needle	  was	  used	
  to	  inject	  the	  contrast	  agent.	
  Prior	  to	  injection,	
  the	
  tail	  vein	  was	  immersed	  in	  warm	  water	  for	  30	  to	  60	  seconds	  to	  increase	  blood	  flow	  to	  the	
  tail	  and	  dilate	  the	  vessels.	
  	  NOTE:	
   Refer	  to	  the	  MediLumine’s	
  Publication	  ‘Optimal	
  Usage	  of	  Fenestra	  Nanoemulsions’	  for	  recommendations	  and	  detailed	
  instructions	  related	  to	  dosage,	  animal	
  preparation,	  and	  administration	  at	  http://www.medilumine.com/resources/user-­‐guides.	  	  	  	  	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  Image	
  Acquisition	  EquipmentImTek	  MicroCAT	  II	  microCT	  scanner.	  

Animal PlacementAnesthetized	  mice	  were	  placed	  on	  the	  imaging	
  table	  in	  the	  prone	  position.	  Use	  of	  gas	  anesthesia	  and	  its	  associated	  tubing	  for	  gas	  supply	  required	  the	  animals	  to	  be	  positioned	  with	  their	  tails	  toward	  the	  gantry.	  
The	  desired	  body	  region	  was	  selected	
  from	  the	  scout	  view	  as	  the	  anatomic	  landmark	
  for	  image	  acquisition.	  

SettingsThe	  settings	  selected	
  for	  this	  medium	  resolution	  contrast-­‐enhanced	  study	  were	  as	  follows:	  X-­‐Ray CameraParameterSetting

Serial	  CCD	
  Length2048Parallel	  CCD	  Length3072

Serial	  Bin	  Factor2Parallel	
  Bin	  Factor2Exposure	  Time750	  msWarp	  CorrectionYes

Defect	  Map	  Correction

NoParameter	  SettingX-­‐Ray	  Voltage80.0	  kVpAnode	  Current500.0	  AX-­‐Ray Tube	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  CT ScanParameterSettingRotating	  Stage	  Start	  Position0.000	
  degreesBed	  Axis	  Position

313.863	  mmBed	  Height43.026	
  mmDetector	  Position0.000	  mmTotal	  Rotation

360	  degrees

Number	  of	  Rotation	  Steps520

Number	  of	
  Axial	
  Bed	  Steps0

Number	  of	  Detector	  Steps0

Number	  of	  Acquired	
  Calibration	
  Projection	  Display	  Period
Exposures
Raw	  Data20

Real	  Time	  ReconstructionNo

Total	
  Scan	
  Time11.07	  min1
Written	  to	  FileScanner Geometry SetupParameterSetting

Source	  to	  Detector	  Distance309.600	  mmSource	  to	  Center	  Distance

256.200	
  mmPhysical	  Detector	  Pitch32.700	
  mmDetector	  Array	
  Height2048	  elementsDetector	  Array	
  Width3072	  elementsCenter	
  Offset4.7	  unbinned	
  detectors	  Images	  were	  obtained	
  immediately	  after	  administration	  (T	  =	  0)	  with	  subsequent	  scans	  acquired	  at	  15,	
  30,	
  60,	  120,	
  180,	  and	  240	  minutes	  post-­‐injection.	  Beginning	
  at	
  T	  =	  0,	  vascular	  contrast	  rapidly	  increased	  to	  a	  level	
  that	  was	  sustained	  at	  all	  time	  points	  for	  the	  remainder	  of	  the	  study.	
  The	  liver	
  showed	
  a	  slight	  increase	  in	  CT	  density	  due	  to	  its	  significant	  vascular	  supply,	  while	  the	  highly	  vascular	  structure	  of	  the	  spleen	  resulted	  in	  a	  high	  level	  of	  contrast	
  enhancement.	  After	  the	  four-­‐hour	  time	  point,	  vascular	  contrast	
  enhancement	  declined	  gradually	  as	  Fenestra	  VC	  underwent	  hepatobiliary	
  elimination.	  

MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  	  Data	  Reconstruction	  and	  Visualization	  Data ReconstructionMachine-­‐based	  reconstruction	
  does	  not	  allow	
  for	  down-­‐sampling	  of	  projections	
  prior	  to	  reconstruction,	
  as	  does	  software-­‐based	  reconstruction	  programs.	  However,	  down-­‐sampling	  was	
  unnecessary	  for	  the	  selected	  resolution	  in	  this	  study.	  

NOTE:	   Reconstructed	  image	
  files	  can	  be	
  stored	  as	  CT	  or	  ATT	  files,	
  which	  can	  be	  exported	  to	  a	  3D	  visualization	
  application	  such	
  as	  Visage	  Imaging’s	  Amira	  software	  for	  viewing	
  as	  axial,	  coronal,	  and	  sagittal	  images,	  in	  addition	  to	  a	  number	
  of	  other	  image	  representations.	  

ParameterSetting

Number	  of	
  Voxels	
  in	  Volume512x512	
  (transaxial)	   768	   (axial)

Voxel	  Size	
  100	  μM	  (transaxial)	  	  150	  μM	  (axial)	  Reconstruction	  Filter

Shepp-­‐Logan

Reconstruction	  AlgorithmFledkamp	  cone-­‐beamData VisualizationData is routinely imported from reconstruction programs as raw CT image data or as bitmaps windowed to avascular contrast setting. Data can be viewed in Amira using the Standard Display format with simultaneousdisplay of the axial, coronal, and sagittal images, or as 3D isosurface images that can be manipulated to viewanatomic structures with or without orthoslice display of 1, 2, or all 3 of the planar slices. The isosurface imagecan also be cropped to eliminate extraneous data and saved as an Amira map file, which can accelerateisosurface viewing and save storage space.

Representative	  Images	  Representative	  images	  from	  studies	  conducted	  in	  CT-­‐26	  tumor-­‐bearing	  BALB/c	
  male	  mice	  using	  Fenestra	
  LC,	
  and	  that	  were	  obtained	  with	  MicroCAT	  II	  scanner,	  are	  provided	  in	  the	  figures	  below.	  	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  	  	  	  	  	  	  	  	  	  	  	  	  Figure	  1.	  Non-­‐contrast	  coronal	  scan	  of	  a	  male	  mouse.	
  Poor	  soft	  tissue	  contrast	  is	  evident	  in	  the	  thoracic	  and	  abdominal	  cavities.	  The	  bright	  spots	  observed	  in	  the	  intestines	  were	
  caused	  by	  minerals	  in	  the	  rodent	  chow	  that	  attenuated	  X-­‐ray	  energy.	  

	  Figures	  2.	  Axial,	  coronal,	  and	  sagittal	
  views	  of	  a	  male	
  mouse	  15	  minutes	
  after	  IV	  injection	  of	  Fenestra	  VC.	  a.	
  The	  axial	  image	  depicts	  both	  ventricles	  of	  the	  heart,	  as	  well	  as	  several	  major	  vessels	  in	  the	  thoracic	  cavity.	  b.	  The	  coronal	  image	  shows	  the	  inferior	  vena	  cava,	
  the	  renal	  veins,	  additional	  renal	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  vasculature,	  as	  well	  as	  several	  veins	
  lower	  in	  the	  abdomen.	  The	  spleen	  (seen	  adjacent	  to	  the	  left	  kidney)	  is	  slightly	  enhanced	  due	  to	  its	  high	  degree	  of	  vascularity.	  c.	  In	  the	  sagittal	
  view,	  the	  inferior	
  vena	  cava	  and	  a	  short	  segment	  of	  the	  portal	
  vein	  can	  be	  seen	
  below	  the	  dome	  of	  the	  liver,	
  while	  the	  heart,	  aorta	  and	  several	  other	  vessels	  can	  be	  seen	  in	  the	  thoracic	  region.	  	  	  Figures	  3.	  Coronal	  and	  sagittal	
  scans	  of	  a	  male	  mouse	  obtained	  3	  hr	  after	  IV	  administration	  of	  Fenestra	  VC.	  a.	  Coronal	  image	  shows	  Fenestra	  VC	  continued	  to	  produce	  vascular	  contrast	  3	  hr	  after	  injection,	  clearly	
  delineating	  the	  ventricles	  and	
  ventricular	
  wall,	  the	  aorta,	  and	  the	  carotid	  arteries	  in	  the	  neck.	  
Small	  vessels	  are	  clearly	  observed	  in	  the	  liver,	
  which	  is	  only	  slightly	  enhanced	  due	  to	  the	  initial	
  elimination	  of	  the	  contrast	  agent,	  although	
  the	  gall	  bladder	  has	  yet	  to	  enhance	  as	  a	  result	  of	  excretion	  of	  the	  metabolized	
  agent.	  b.	  This	  sagittal	  view	  shows	  	  	  a	  major	  vessel	  diagonally	  traversing	
  the	  liver	  with	  a	  number	  of	  smaller	  vessels	  seen	
  in	  cross	  section.	
  The	
  heart	  and	
  the	
  aorta	  are	  readily	
  visualized,	  as	  is	  another	  vessel,	  which	  appears	  to	  bifurcate	  just	
  below	  the	  base	  of	  the	  skull.	  c.	  Sagittal	  image	
  shows	  the	  descending	
  aorta	  and	  several	  other	  arteries	  in	  the	  abdomen.	  The	  heart,	  aorta,	  and	  several	  additional	  vessels	
  are	  seen	  in	  the	  thorax	
  and	  neck	  of	  the	  mouse.	  Chow-­‐induced	  artifacts	  are	  evident	  in	  the	  intestines	  in	  all	  three	  images.	  

MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  	  Figures	  4.	  Volumetric	  representations	  of	  vascular	
  enhancement	  from	
  scans	  of	  a	  male	  mouse	  3	  hr	  after	  IV	  administration	  of	  Fenestra	  VC.	  Fenestra	  VC	  produces	  prolonged	  vascular	  contrast	
  enhancement	
  throughout	  the	  entire	
  body	  from	  a	  single	  peripheral	  injection.	  	  	  	  	  	  	  	  	  	  	  	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  
The	  custom	  parameters	  selected	
  for	  this	  study	  were	  as	  follows:	  	  	  ParameterSettingNumber	
  of	
  Samples512

Number	  of	  Projections3072Integration	
  Time200	  
You	  should	
  note	  that	  changes	
  in	  these	  parameters	  can	  influence	  both	  total	  image	
  acquisition	
  time	  and	  image	  quality.	  The	  average	
  time	  for	  image	
  acquisition	  for	  vascular	  imaging	  with	  Fenestra	  VC	  was	  approximately	  eight	  minutes	  for	  acquiring	
  the	  scout	
  view	  and	  the	  250	  step	  study	
  with	  the	  vivaCT	  40	  scanner.	  To	  save	
  time,	  you	  can	  “pre-­‐calibrate”	  the	  scanner	  so	  that	  the	  scans	
  begin	  as	  soon	  as	  the	  animal	  is	  inserted.	  
Images	  were	  obtained	
  immediately	  after	  administration,	
  T=0,	  with	  subsequent	  scans	  acquired	  at	  several	  time	  points.	  Beginning	  at	  T=0,	  vascular	  contrast	  rapidly	  increased	  to	  a	  level	
  that	  was	  sustained	  for	  the	  remainder	  of	  the	  study.	
  The	  liver	
  showed	
  a	  slight	  increase	  in	  CT	  density	  due	  to	  its	  significant	  vascular	  supply,	
  while	  the	  highly	  vascular	  structure	  of	  the	  spleen	  resulted	  in	  a	  high	  level	  of	  contrast	
  enhancement	
  in	  that	  organ.	  Beyond	  the	  6-­‐hour	  time	  point,	  vascular	  contrast	
  enhancement	  declined	  gradually	  as	  Fenestra	
  VC	
  began	  to	  undergo	  hepatobiliary	
  elimination.	  You	  should	
  note	  that	  the	  gray	
  scale	  may	  have	  to	  be	  adjusted	  to	  display	  low	  contrast	  regions.	  

Data	  Reconstruction	  and	  Visualization	  Data ReconstructionParameter	  Setting

Number	  of	
  Voxels	  in	  Volume1024	  x	  1024	  or	  User-­‐defined	  values	  (Custom	  Protocol)

Voxel	  Size

FOV	  divided	  by	  the	  number	  of	  samples

Reconstruction	  Filter	  	  	  Reconstruction	  Algorithm	
  	  

None	
  Cone-­‐beam	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  Data VisualizationAcquired	  images	  can	  be	  visualized	  in	  the	  evaluation	  window.	  You	  should	
  note	  that	  low	  contrast	  images	  may	
  be	  viewed	  better	  with	  a	  black	  background	
  than	  the	  standard	
  blue.	  You	  can	  toggle	  between	
  backgrounds	  by	  clicking	  on	  the	  scalar	  on	  the	  right	  side	  of	  the	
  window	  or	  by	  choosing	  the	  LUT	  option	  and	  then	  selecting	  the	  correct	  LUT	  and	  optimizing	  the	  contrast.	  
In	  addition	  to	  exporting	  AIM	  and	  ISQ	  files,	  images	
  can	  also	  be	  exported	  as	  TIFF	  files.	  You	  should	  note	
  that	  AIM	
  files	  can	  be	  converted	  to	  the	  DICOM	  file	  format	  for	  export.	
  Any	  data	  visualization	  program	  capable	  of	  reading	  and	  displaying	  TIFF	
  or	  DICOM	  file	  formats	  can	  be	  used	  to	  display	
  the	
  data.	  	  Representative	  Images	  	  Representative	  images	  from	  studies	  conducted	
  in	  normal	
  mice	  with	  Fenestra	  VC,	  and	
  that	  were	  obtained	  with	  SCANCO	  Medical’s	
  vivaCT	  40	  scanner,	  are	  provided	  in	  the	  figures	  below.	  

Pre-contrast Exam	  Figure	  1.	  Non-­‐contrast	  axial	  scan	  of	  a	  mouse.	
  Poor	  soft	  tissue	  contrast	  is	  evident	  in	  the	  thoracic	  cavity	  and	  heart	  chambers.	  

MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014  	  	  	  Fenestra VC Exam	  Figure	
  2.	  Axial	
  views	  of	  a	  mouse	  2	  hours	
  after	  IV	  injection	  of	  Fenestra	  VC.	
  Images	  a	  and	
  b	  depict	  both	
  ventricles	  of	  the	  heart,	  as	  well	  as	  several	  major	  vessels	  in	  the	  thoracic	  cavity.	  Image	  c	  also	  shows	  both	  cardiac	  chambers,	  as	  well	  as	  additional	  thoracic	
  vasculature	
  near	  the	  bottom	  of	  the	  heart.	  	  Figure	  3.	  Coronal	  view	  of	  a	  mouse	  2	  hours	
  after	  IV	  injection	  of	  Fenestra	  VC.	
  The	  image	  shows	  both	  cardiac	  chambers,	  as	  well	
  as	  small	  hepatic	  vessels	  	  	  MediLumine  Inc.    

medilumine.com     5795  De  Gaspe  Avenue,  Suite  #112  CTS  Healthcare  Campus  Montreal  (Quebec),  Canada  H2S  2X3  	      	  Tel:  514.360.1574    Toll  free:  1.844.360.1574    Cell:  514.929.9744  Fax:  514.360.6014