Intravital Imaging

Headed by Prof. Colinda Scheele

E-mail: colinda.scheele@kuleuven.be
Location:  08.152, O&N4, Campus Gasthuisberg                  
ORCID: 0000-0001-8999-5451

Tumors are highly dynamic ecosystems, in which the tumor cells and their micro-environment consisting of many different cell types, continuously evolve over time. To understand this intricate and complex interplay, it is crucial to study tumor evolution in space and over time, i.e. in 4 dimensions. With intravital microscopy single cells can be followed over multiple days to weeks in unperturbed in vivo tissues which makes intravital imaging the only method to study the phenotypic behavior and plasticity of individual tumor cells and wild-type cells in their natural niches. This technology has further been applied to study metabolic, neurologic, immunological interactions and infectious diseases.

The CCB Intravital Imaging Expertise Center harbors two high-end multiphoton microscopy systems dedicated to the in vivo imaging of cells and their native micro-environment. To this end a wide array of fluorophores, biosensors and in vivo imaging dyes can be supplied and recommended by the center. The setup of the imaging systems is specifically designed to allow deep in vivo tissue imaging in zebrafish and of both ventrally and dorsally localized tissues of interest in living mice. Both microscopes are specifically equipped with several accessories for long-term, multiday imaging sessions to follow tumor cell behavior over longer time periods. To gain optical access to the tissues of interest, small imaging windows have been developed and are utilized to visualize individual tumor cells in their native environment.

Services

• Design, planning and execution of intravital experiments
  • Choosing and optimizing biosensor and/or dyes and stains from our collection
  • Aiding in writing and review ECD applications
  • Training on 2-photon microscope use
  • Performing and training of surgical techniques (e.g. optical windows for various target organs, including but not limited to subcutaneous tumours, mammary gland/tumours, pancreas, liver, spleen, lung and brain)
• Tissue imaging ex vivo
  • Customized tissue clearing and staining of intact tissue
  • Single photon and 2-photon imaging of stains, tissue autofluorescence (e.g. SHG, collagen I)
  • Tissue size can be in the cm range
• Post imaging processing
  • 3D rendering of cleared tissues (e.g. tumours, whole organs)
  • Quantification e.g. blood flow, extracellular matrix (ECM) content and organization
  • Motion correction of intravital imaging

In house systems

Leica SP8 

Location: O&N4 08.149

• upright system with heated chamber
• Single photon laser lines: 405, 458, 476, 488, 496, 514, 561, 633
• 4 internal HyD and 1 PMT detectors
• 2-photon lasers:
  • 680-1060 tunable wavelength for 2P
  • tunable OPO 1080-1300
• 4 external HyD detectors
• available filter cubes:
  1. 460/50, 525/50, dichroic 495
  2. 610/25, 736/128, dichroic 790
  3. 525/50, 585/40, dichroic 560
  4. 624/40, 685/40, dichroic 650

Leica SP8 DIVE             Location: O&N4 09.339

• Inverted system with heated chamber
• connected anesthesia unit
• Single photon laser lines: 405, 488, 552, 638
• 2 internal HyD and 1 PMT detectors
• 2-photon lasers:
  • 1040 (fixed wavelength 2P)
  • 680-1060 tunable wavelength for 2P
• 4 tunable external detectors (3 HyD, 1 PMT)

The CCB Intravital Microscopy Expertise Center strives to stay at the forefront of technology development. We continuously develop and invest in improved microscopy technologies, new imaging tools and refined surgeries for 3D and 4D in vivo imaging.