Imaging Unit
Imaging Transgene Expression
in Live Animals
Monitoring the expression of therapeutic genes in targeted tissues in disease models is of utmost importance for assessing the effectiveness of gene therapy delivery systems. We applied a new light detection cooled CCD camera (CCCD) and/or a lightool camera (CCD) for GFP and RFP expression, for in-vivo assessment of commonly used gene therapy delivery systems such as ex-vivo manipulated cells, viral vectors and naked DNA, continuously, without the need to sacrifice animals.
We examined a variety of criteria related to real time monitoring of luciferase (luc), or green protein fluorescence (GFP) or red protein fluorescence (RFP) gene expression in a wide spectrum of tissue including bone, muscle, salivary glands, dermis, liver, peritoneum, testis, teeth, prostate and bladder in living mice and rats.
These criteria included determination of the efficiency of infection/transfection of various viral and non-viral, delivery systems, promoter specificity and the visualization, as well as the ability of luciferin or GFP or RFP to reach various organs. In conclusion, we showed that the cooled CCD photon detection system and the CCD GFP or RFP detection systems are simple, reproducible and applicable methods that enable the continuous monitoring gene delivery system in living animals.
Cell-viZio is a unique imaging system for molecular imaging
of live animals at the cellular level
· Molecular imaging at the cellular level
· Fluorescence optical imaging
· Fiber-based system
· Real time
· Longitudinal Studies
Technical features
· Lateral resolution down to 2.5 microns
· Axial resolution 15-20 microns
· 488 nm excitation, 500 to 650 nm collection
· 12 frames/second
· Minimally invasive
Interested investigators are requested to download and complete the online application ImagingUnit order form. and hand deliver or e-mail it to:
Evelyne Zeira
The Goldyne Savad Institute of GeneTherapy
Hadassah UniversityHospital,
Ein Jerusalem, 91120,Israel
Phone: 02-677-8782(lab)cell.phone:73640
Fax:02-643-0982
e-mail:evezeira@hadassah.org.il
or
Hagit Yotvat
The Goldyne Savad Institute of GeneTherapy
Hadassah UniversityHospital, Ein Kerem
Jerusalem, 91120,Israel
Phone: 02-677-8881(lab)
Fax:02-643-0982
The researcher has the following obligations:
He/she should bring:
- The Luciferin substract(Beit Haemek-Promega-cat# E6103). The 250 mg vialshould be diluted with 40 ml of sterile PBS which will give 80 pictures. Eachmouse is injected with Luciferin at 126 mg/kg or 3.15 mg/mouse in a total volumeof 0.5 ml (from the dilution of 40 ml take 0.5 ml permouse).
- Syringes, needles, anestheticmaterials.
- The researcher will be responsible forinjecting anesthetic solutions, Luciferin, and explain to Hagit, our technician,which side of the mouse needs to be pictured.
- It is recommended that researchers requiringto use any of our systems should first contact Evelyne Zeira or HagitYodvat.
- All mice used for this purpose need to be SPFmice and all new projects will have to be discussed first with Evelyne Zeira orDr. Yfat Uzi, the veterinarian.
- A copy of this form will have to be submittedto Dr. Yfat Uzi at the SPF Animal Facility, before starting any experiment, andhave this form signed. A copy of the signed form should then be brought to Evelyne Zeira or Hagit Yodvat.
- The researcher is requested to provide a shortdescription of the project on a separate page.
Important: We would appreciate, that any publication arising from the use of imaging technologyacknowledge this service as follows:
“The imaging technology services were developed at the Israeli National Knowledge Center for GeneTherapy of the Israeli Ministry of Science, the Goldyne Savad Institute of Gene Therapy, Hadassah University Hospital, Jerusalem, Israel.”