Functional Immune Imaging

PI/Head: Lai Guan NG, Ph.D.

Manager: Akhila BALACHANDER, Ph.D.

The SIgN Functional Immune Imaging Platform applies multi-photon microscopy on intact living tissues, allowing the in vivo dynamics of immune-cell behaviour to be examined in real-time

Background

Analyses of the in vivo dynamics of immune-cell behaviour can provide knowledge as to how orchestrated cell migration and cellular interactions may contribute to physiological and pathological outcomes during immune responses.  Dr Lai Guan Ng’s research group has developed several imaging models that are available to all SIgN researchers via the Functional Immune Imaging Platform. These imaging models can be used to study the dynamics of immune-cell migration and cellular interactions within intact mouse tissues. Examples of established intra-vital imaging models include bone marrow, brain, skin, lung, spleen, kidney and muscle. 

Technologies and Approach
A. Multi-Photon Microscopy

The Functional Immune Imaging Platform, as part of the Advanced Microscopy Cluster (AMC), houses a series of advanced microscopy modalities for in vivo imaging.

Our equipment includes:
•    Upright modular 2-photon microscope (LaVision Biotec)
•    Dual head 2-photon microscope for both upright and inverted applications (LaVision Biotec)
•    High speed and high resolution 2-photon microscope (Olympus)

B. Light Sheet Microscopy 

The platform also houses an Ultramicroscope, a light sheet illumination microscope for optically cleared whole organ/tissue. This technique allows 3D volumetric macroscopic imaging with high resolution.

C. Confocal Microscopy 

The Confocal Microscopy combines cutting edge equipment with dedicated in-house expertise to facilitate research at SIgN. It has facilities for extended live-cell imaging under controlled temperature and CO2 conditions, 3-D imaging, FRET (Fluorescence Resonance Energy Transfer), FRAP (Fluorescence Recovery After Photobleaching), Spectral deconvolution, Photo-switchable fluorophores and Image processing and analysis. On site consultation is available for experimental planning, labelling methodologies and image acquisition. We also conduct periodic training courses appropriate for a range of skill levels.

Our equipment includes:
•    FV-1000 confocal system comprising an inverted Olympus IX81 microscope, FV1000 scan head, 405, 488, 515, 559, 635 nm laser lines, 4 PMTs for fluorescence detection, one PMT for transmitted light detection, 10x, 20x, 40x, 60x Water, 60x Oil, 100x Oil lenses all equipped with DIC prisms
•    Offline workstations for Fluoview, TissueQuest and HistoQuest
•    Olympus IX81 widefield fluorescence microscope
•    Zeiss Axiovert 200 widefield fluorescence microscope
•    Nikon Eclipse TS100