Widefield Microscopy
MNI Widefield Microscopy Facility
Welcome to the MNI Widefield Facility. This is a jointly-operated facilitiy between the Heit, Dikeakos and Kerfoot labs which provides users with access microscopes, analysis computers, and a cryostat for sample preparation. All users must be registered and trained prior to using any equipment in the facility. A full description of the available equipment and its capabilities can be found below.
Training & Fees
MNI Widefield Facility fee & training schedule (PDF, August 2019)
Widefield Microscope Manual (PDF, April 2014)
Widefield Microscope
The widefield microscope is a custom-designed Leica DMI6000 system intended for high and super-resolution imaging of living cells. This system features:
- 40x/1.30 NA, 63x/1.40 NA and 100x/1.40NA objectives.
- A heated live-cell stage with humidified air/CO2 perfusion.
- Adaptive focus control and autofocus capabilities.
- A high sensitivity EM-CCD camera capable of frame rates of up to 50 FPS.
- A large field of view CMOS camera.
- Tiling and point-visiting capabilities.
- Chroma Sedat Quad rapid filter set. 50 ms switching times between channels, with filters for DAPI, FITC, Cy3 and Cy5
- Secondary filter set for FITC ratiometric imaging and CFP/YFP FRET.
- DIC white light imaging
- Capable of multi-colour SRRF super-resolution microscopy
Tissue Imaging Microscope
The tissue imaging microscope is built for generating large, tiled immunofluorescence, immunocytochemistry, and stained images of tissue sections. It features:
- A range of low- and medium-magnification lenses
- DAPI, FITC, Cy3 and Cy5 channels
- 16-bit and colour CMOS cameras
- Advanced tiling and zoom capabilities
Cryostat
We have a Leica CM18602 cryostat for cutting frozen sections as part of the preparation process for immunostaining or immunohistochemistry.
Deconvolution Computer
This is a dedicated 2-chip Xeon workstation for performing deconvolution and 3D rendering of images acquired on the DMI6000 widefield system using Leica LAS-X software.
Note: access to this resource is limited during Westerns COVID-19 limited work capacity. Contact Dr. Heit if you require access to this resource.
Analysis Computer (Tux)
This is a high-powered analysis workstation built for general image processing tasks. This station runs Ubuntu Linux and has the following software packages installed:
Image Processing:
- FIJI & ImageJ
- GIMP and Inkscape
- Icy
- ChimeraX
- CellProfiler
- Ilastik
- Matlab + our MIiSR and Particle Tracking [1, 2] software
Imaris Workstation
This is a high-powered workstation for performing image analysis using Imaris software.
Note: access to this resource is limited during Westerns COVID-19 limited work capacity. Contact Dr. Heit if you require access to this resource.
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