Day One Topics: Basic Instrument Operation - Image Acquisition, Imaging Storage, Data Analysis
System Set-Up
IVIS Cleaning Procedure & Identification of Proper Cleaners
Using imaging parameters of IVIS System Control Panel (time, binning, f-stop, FOV level) in order to acquire a bioluminescent image using non-biological phantoms
Properly labeling the Image Info Panel
Saving Acquired Images
Theories for data analysis, drawing proper Regions of Interest (ROI’s), importance of using correct units
Special ROI tools
Exporting data
Other options using the measurement table
Browser Features: Recalling saved images, sorting images in browser, Load as a Sequence function
Features for Advanced & More Efficient Data Analysis
Introduction to Fluorescent imaging: Brief discussion of instrument background and tissue auto fluorescence
Acquire fluorescent images using non-biological phantoms practicing techniques for instrument and auto fluorescent subtraction
Printing Single or Multiple Images
Displaying Images for Presentation or Publication
Day Two Topics: Animal Models -Practical aspects of luciferase-based bioluminescent
Presentation: Oncology, Metabolic Disease, Inflammation, and Infectious Disease Models
Light Producing Transgenic Animals and their proper use
Animal Preparation
Sedation of Animals, use of Induction Box, Anesthesia while Imaging
Isoflurane Anesthesia System
General procedures for luciferin preparation, storage and use
Biology of luciferase-based imaging
Principles of light emission by the firefly luciferase
Key factors for consistent signals in bioluminescent imaging
Pharmacokinetics of D-luciferin in rodents and its impact on imaging
D-luficerin – preparation, storage and delivery to animals
Image acquisitions
Optimizing imaging parameters for a single image
Image analysis and presentation
Comparing images from longitudinal experiments
Proper design of experiments
Practical aspects of imaging large cohorts of animals
LPTAs: b -actin example
How to address user’s particular scientific questions by using luciferase-based bioluminescence imaging
Recommendations for use of our biological products
Models with grafted luciferase – labeled biological entities (mammalian cells, or bacteria)
LPTAs
Discussion: Strengths/Features of bioluminescent imaging
Day Three Topics: Fluorescence Imaging & 3D Bioluminescence Imaging
Presentation: Fluorescence imaging: Principle and Biological Applications
Tissue auto fluorescence and imaging considerations
Fluorescence imaging reagents: fluorophores, quantum dot, and fluorescent protein
Selection of imaging filters for excitation and emission
Background fluorescence and its impact on sensitivity and reliability of fluorescence signal detection
Possible ways to remove fluorescence background
Auto fluorescence background
Instrument background
Epi illumination and its advantages
How to address user’s particular scientific questions by using fluorescence imaging
Data quantification, processing and presentation
All the trainees will be required to set up the imaging sequence, acquire the images, and perform data quantification
Presentation: Principles of 3D Bioluminescent Imaging
Advantages over ROI measurements
Methods
Luciferase emission and tissue optical property variation with light wavelength
Determination of animal surface for tissue-air interface
Selection of imaging parameters for 3D Bioluminescence Imaging
Using narrow-bandpass spectral filters
Data required for surface rendering
Consideration of Luciferin kinetics for image sequences
Data analysis of 3D Bioluminescent Images
Demonstration of 3D reconstruction
Source quantification and tumor depth measurement
Working with the Mouse Atlas
Presenting 3D representative images
IVIS - Lumina Series, Ivis 50, 100, 200, 3D, Spectrum - Note*** Please refer to course description to determine the amount of days your specific training will require.
