The Live Cell Imaging Facility (LCIF)
Cutting-edge microscopy and image analysis services, available to researchers at UM as well as those in academia and industry
The Live Cell Imaging Facility offers advanced microscopy technology for capturing high-quality images of both live and fixed cells. From experiment design to image analysis, we provide technical support to help you achieve your imaging objectives. Explore our website to learn more about our services and capabilities. Our users investigate a variety of topics including genetic modifiers of neurological conditions, synaptic changes in neurodevelopmental and neurodegenerative disorders, neuron-glia interactions, glia-regulated neurovascular coupling, and brain and spinal cord injury mechanism and repair. To address such questions, our users take advantage of our advanced imaging modalities to assess dendritic length and branching, synaptic counts, synapse component in cell culture and brain slice, microglia morphology, Ca2+ signaling in astrocytes, neurons, pericytes, and endothelia cells, and changes in blood vessel diameter and blood flow.
Hours of usage: 7771 hours
Accessed by: 60 staff/trainees
OUR SERVICES
Microscopy
We provide consultation and training services for our range of microscopes, as well as the option to conduct experiments on your behalf.
Confocal
Confocal microscopy is an advanced optical imaging technique that uses fluorescence to generate high-resolution, high-contrast images with improved axial resolution compared to traditional epi-fluorescence. The technique involves illuminating the specimen with a laser excitation source at a single location (corresponding to a single pixel in a digital image) and detecting the emission through a pinhole located in a conjugate focal plane. The laser is then scanned over the entire field of view to record fluorescence emission for a small axial volume at each location. Our confocal microscope offers down to 120nm resolution and is equipped with incubation chambers for live cell imaging over long periods, allowing for a diverse range of biological specimens to be imaged.
Multiphoton
Unlike confocal microscopy which uses a pinhole to reject out-of-focus light, multi-photon microscopy utilizes a pulsed infrared laser to excite fluorescence only within the diffraction-limited spot of the objective lens, creating an inherent optical section. This has two main advantages. Firstly, the longer wavelength of the near-infrared laser allows it to penetrate biological tissue more efficiently, making it possible to image thicker samples than with traditional confocal microscopy. Secondly, the inherent nature of the optical section generated by photon crowding enables direct fluorescence detection without having to travel back through a scanning system, improving the ability to capture scattered light, which is more prevalent in thicker samples. Multi-photon microscopy is particularly effective for imaging thick and opaque samples, such as tissue sections several hundred microns thick or even the intact brain of a live mouse.
Laser Microdissection
Laser capture microdissection (LCM) is a precise technique for isolating cells from their surrounding tissues under direct microscopic visualization, using a laser beam. This method allows specific cells to be selectively removed, separating them from unwanted cells to obtain enriched cell populations. These isolated cells can be utilized to generate cDNA libraries, conduct transcriptome or proteome analyses, or for other research purposes that require pure cell populations.
Analysis
We provide consultation and training services for the most appropriate analysis for you, as well as the option to conduct experiments on your behalf.
On your computer
- FIJI ImageJ: standard image processing including area measurement, object counts, colocalization, and more.
- Cellular and Hemodynamic Image Processing Suite (MATLAB): Analysis of functional images of cells and blood vessels including calcium analysis and hemodynamic measurements.
Remote or in-lab analysis
- IMARIS: 3D rendering and analysis: Deconvolution, Filament tracing, colocalization, and more.
- Huygens: WF, Confocal, Multi-photon, and array detector deconvolution, drift and motion corrector, chromatic aberration corrector, crosstalk corrector, quality control, and more.
Carl Zeiss LSM 880 with AiryScan detector
Capabilities
- Super-resolution imaging with AiryScan (1.7X improvement over diffraction-limited imaging)
- Multichannel imaging
- Spectral imaging and unmixing
- Live cell imaging
- FRET, FRAP, FLIP photo-activation/-conversionZ-stack, tile scanning, time lapse
Microscope
- Zeiss Axio Observer Z1
- Motorized XY scanning stage
- Motorized focus control (Z) with Definite Focus
- Stage Top Chamber with Environmental Support (temperature, humidity and CO2)
Objectives
- 5x, 0.13 NA
- 20x, 0.8 NA
- 40x, 1.2 NA, Water Corr
- 63x, 1.3 NA, Oil
Illumination
- Transmitted Light Illuminator (Halogen lamp)
- Epi Fluorescence Illuminator (HXP 120V; widefield visual observation)
- available laser lines: 405nm, multiline argon (458nm, 488nm, 514nm), 561nm, 633nm
Detectors
- Two Conventional PMTs
- One high sensitivity GaAsP detector
- One AiryScan Super Resolution 32-channel GaAsP Detector
Bruker Ultima In Vitro
Capabilities
- Two-photon excitation with dual channel detection allows for deep tissue penetration and imaging of GFP together with mCherry/TdTomato.
- Multiarea (XYZ), time-lapse imaging with environment control (solution perfusion, O2/CO2, temperature) for live imaging in tissue sections
- Photostimulation and uncaging
Microscope
- Olympus BX61WI upright microscope with motorized focus control (Z drive)
- Fully motorized stage for precise repeatable movement (XY) of the specimen with respect to the fixed microscope
- Stage top chamber with environmental support (temperature, humidity and C2)
Objectives
- 10x, 0.3 NA water immersion
- 40x, 0.8 NA water immersion
Illumination
- Transmitted light illuminator (Halogen lamp)
- Ti:S laser tuneable between 690 to 1040 nm
Detectors
- Two photomultiplier detector pairs (non-descanned, above stage and substage): for high sensitivity capture of emitted fluorescence (both reflected and transmitted).
- Equipped with emission filters for two channel acquisition of GFP and mCherry/TdTomato.
- Transmitted light detector with Dodt gradient contrast
Bruker Ultima In Vivo
Capabilities
- Two-photon excitation with dual channel detection allows deep tissue penetration and fluorescence imaging.
- The system is adaptable, with optional stage and specimen chamber available for imaging from tissue sections, but is primarily designed for imaging from live rodents, for example via a cranial window (thinned skull or open-skull glass window) at depths of up to 1 mm from the brain surface
- Capable of live imaging at video frame paired with simultaneous photo-stimulation.
Microscope
- Olympus BX61WI upright microscope with motorized focus control (Z drive)
- Fully motorized stage for precise repeatable movement (XY) of the specimen with respect to the fixed microscope is available.Stage top chamber with environmental support (temperature, humidity and O2/CO2) is available for use.
Objectives
- 10x, 0.3 NA water immersion
- 20x, 1.0 NA water immersion
- 40x, 0.8 NA water immersion
Illumination
- Transmitted light illuminator (Halogen lamp)
- Fast resonance scanner with inputs for two Ti:S lasers tuneable between 690 to 1040 nm, one of which is dedicated for photostimulation
Detectors
- Two high-sensitivity GaAsP detectors, configured to capture emitted fluorescence from up to three distinct fluorophores (e.g., blue, green or red fluorescent emitters) in real-time.
- Transmitted light detector with Dodt gradient contrast
Carl Zeiss PALM MicroBeam
Capabilities
- Microbeam laser unit (pulsed 355nm) allowing laser cutting and non-contact capture of both fixed and live samples
- Multichannel imaging
- Live cell imaging
- Timelapse, time series, timed capture
Microscope
- Zeiss Axio Observer Z1
- Motorized XY scanning stage and focus control (Z)Stage top chamber with environmental support (temperature, humidity and CO2)
Objectives
- Fluar 5x, 0.25 NA
- Fluar 10x, 0.25 NA
- LD Plan-Neofluar 20x, 0.4 NA CorrLD
- Plan-Neofluar 40x, 0.6 NA Corr
Illumination
- Transmitted light illuminator (Halogen lamp)
- Epi fluorescence illuminator (HXP 120V)
- Filter based excitation/emission for DAPI, GFP, mRFP, Cy5
Cameras
- AxioCam ICc 5 – colour camera
- AxioCam MRm – monochrome camera
access + Acknowledgement
You can book directly through our Rady Faculty of Health Sciences page on the University of Manitoba website. There you will find all information including consultation, reservations, training, fees, and billing.
We take great pride in assisting you in your scientific endeavors and hope that our services have been helpful in advancing your research goals. We kindly remind you that acknowledging LCIF in your publications is not only a professional courtesy, but also an important factor in securing continued support for our facility.
Here is a suggestion for acknowledging LCIF in your publications:
“Images presented in this publication were collected and/or processed/analyzed at the Live Cell Imaging Facility at the University of Manitoba.”
CONTACT
Noushin Ahmadpour
Imaging Specialist PrairieNeuro Research Center
Kleysen Institute for Advanced Medicine
Department of Pharmacology & Therapeutics
Rady Faculty of Health Sciences
University of Manitoba
SR426, 710 William Avenue
Winnipeg, MB, Canada R3E 0Z3
Dr. Michael F. Jackson
Principal investigator, Facility Director
PrairieNeuro Research Center
Kleysen Institute for Advanced Medicine
Associate Professor
Department of Pharmacology & Therapeutics
Rady Faculty of Health Sciences
SR426, 710 William Avenue
Winnipeg, MB, R3E 0Z3
Publications
Jessica Meza-Resillas, Noushin Ahmadpour, Michael Stobart, Jillian Stobart. Brain Pericyte Calcium and Hemodynamic Imaging in Transgenic Mice In Vivo. J Vis Exp. 2021 Nov 20;(177). doi: 10.3791/62725. PMID: 34866618.
Chetan S. Patil, Hongbin Li, Natalie E. Lavine, Ruoyang Shi, Ankur Bodalia, Tabrez J. Siddiqui, and Michael F. Jackson. “ER-Resident STIM1/2 Couples Ca2+ Entry by NMDA Receptors to Pannexin-1 Activation.” Proceedings of the National Academy of Sciences of the United States of America 119, no. 36 (September 6, 2022): e2112870119. doi: 10.1073/pnas.2112870119.
Noushin Ahmadpour*, Meher Kantroo*, Michael J. Stobart, et al. Cortical astrocyte N-methyl-D-aspartate receptors influence whisker barrel activity and sensory discrimination in mice. Nat Commun. 2024 Feb 21;15(1):1571. doi: 10.1038/s41467-024-45989-3.
Jessica Meza-Resillas, Finnegan O’Hara, Syed Kaushik, Michael Stobart, Noushin Ahmadpour, Meher Kantroo, Shahin Shabanipour, John Del Rosario, Megan C. Rodriguez, Dmytro Koval, Chaim Glück, Bruno Weber, Jillian Stobart. L-Type Ca2+ channels and TRPC3 channels shape brain pericyte Ca2+ signaling and hemodynamics throughout the arteriole to capillary network in vivo. bioRxiv. doi: https://doi.org/10.1101/2024.02.27.582351
FAQ
FAQ Heading
FAQ text
FAQ Heading
FAQ text
FAQ Heading
FAQ text
FAQ Heading
FAQ text
FAQ Heading
FAQ text
FAQ Heading
FAQ text
FAQ Heading
FAQ text
Follow us
Supported by:
Rady Faculty of health sciences
Max Rady College of Medicine
Department of Pharmacology and Therapeutics