Live-Cell Imaging Facility

Location:

• Small Animal and Materials Imaging Core Facility

Capabilities:

• Two-photon excitation with dual channel detection for deep tissue penetration and fluorescence imaging.
• Adaptable system with optional stage and specimen chamber for imaging from tissue sections.
• Primarily designed for live imaging from rodents, using cranial windows (thinned skull or open-skull glass window) up to 1 mm deep 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 and repeatable movement (XY) of the specimen with respect to the fixed microscope.
• Stage top chamber available with environmental support for temperature, humidity, and O2/CO2 control.

Objectives:

• 10x, 0.3 NA water immersion objective.
• 20x, 1.0 NA water immersion objective.
• 40x, 0.8 NA water immersion objective.

Illumination:

• Transmitted light illuminator using a Halogen lamp.
• Fast resonance scanner with inputs for two Ti:S lasers tunable between 690 to 1040 nm, one dedicated for photostimulation.

Detectors:

• Two high sensitivity GaAsP detectors configured to capture emitted fluorescence from up to three different fluorophores (e.g., blue, green, or red) in real-time.
• Transmitted light detector with Dodt gradient contrast.

Location:

• Kleysen Institute for Advanced Medicine, fourth floor

Capabilities:

• Two-photon excitation with dual channel detection 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 capabilities.

Microscope:

• Olympus BX61WI upright microscope with motorized focus control (Z drive).
• Fully motorized stage for precise and repeatable movement (XY) of the specimen with respect to the fixed microscope.
• Stage top chamber with environmental support for temperature, humidity, and CO2 control.

Objectives:

• 10x, 0.3 NA water immersion objective.
• 40x, 0.8 NA water immersion objective.

Illumination:

• Transmitted light illuminator using a Halogen lamp.
• Ti:S laser tunable 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.

Location:

• Kleysen Institute for Advanced Medicine, fourth floor

Capabilities:

• Super resolution imaging with AiryScan, providing a 1.7X improvement over diffraction limited imaging.
• Multichannel imaging capability.
• Spectral imaging and unmixing capability.
• Live cell imaging.
• FRET, FRAP, FLIP photo-activation/conversion techniques.
• Z-stack, tile scanning, and time-lapse imaging.

Microscope:

• Zeiss Axio Observer Z1 microscope.
• Motorized XY scanning stage for precise sample movement.
• Motorized focus control (Z) with Definite Focus feature.
• Stage Top Chamber with environmental support for controlling temperature, humidity, and CO2.

Objectives:

• 5x, 0.13 NA objective.
• 20x, 0.8 NA objective.
• 40x, 1.2 NA Water Corr objective.
• 63x, 1.3 NA objective.

Illumination:

• Transmitted Light Illuminator using a Halogen lamp.
• Epi Fluorescence Illuminator (HXP 120V) for widefield visual observation.
• Laser sources available: 405nm, multiline argon (458nm, 488nm, 514nm), 561nm, 633nm.

Detectors:

• Two Conventional PMTs (Photomultiplier Tubes).
• One high sensitivity GaAsP detector.
• One AiryScan Super Resolution Detector.

Location:

• Kleysen Institute for Advanced Medicine, fourth floor

Capabilities:

• Microbeam laser unit (pulsed 355nm) for laser cutting and non-contact capture of fixed and live samples.
• Multichannel imaging capability.
• Live cell imaging.
• Timelapse, time series, and timed capture features.

Microscope:

• Zeiss Axio Observer Z1 microscope.
• Motorized XY scanning stage and focus control (Z).
• Stage top chamber with environmental support for controlling temperature, humidity, and CO2.

Objectives:

• Fluar 5x, 0.25 NA objective.
• Fluar 10x, 0.25 NA objective.
• LD Plan-Neofluar 20x, 0.4 NA Corr objective.
• LD Plan-Neofluar 40x, 0.6 NA Corr objective.

Illumination:

• Transmitted light illuminator using a Halogen lamp.
• Epi fluorescence illuminator (HXP 120V).
• Filter-based excitation/emission for DAPI, GFP, mRFP, Cy5.

Cameras:

• AxioCam ICc 5 - color camera.
• AxioCam MRm - monochrome camera.

Before starting, check to see that system was left clean and in an operational state. Report any issues to the administrator immediately, as unreported issues will be attributed to the last scheduled user who will receive a warning. Repeat offenders may lose user privileges.

You are responsible for your own data storage. Users should be prepared to save data to an external hard drive or USB flash drive. External drives must be scanned with antivirus software to ensure they are free of virus. Data left on LCIF computers may be deleted at any time without notice in the event local disk space is needed for system operation.

If anything stops working during scheduled use, you must immediately report the issue to the system administrator. Maintenance and repairs are the responsibility of the LCIF. DO NOT attempt to repair or modify any imaging system yourself.

Users will not be billed for repairs under typical circumstances. However, your department may be charged for damage due to negligence or improper handling (for example, if the system was left running unattended over the weekend, a spill causing equipment damage, negligence causing virus or malware infection, etc.)

If a user is booked within the following hour, leave the system on. If not, turn it off. This is to prevent lasers and arc lamps being turned on and off repeatedly. Additionally, lasers should be kept on standby whenever possible to prolong their life.

If imaging is not complete at the end of your booked time-slot, you may continue to work as long as no one is waiting. If another user is booked immediately after, you must finish your task and hand-off the space. Failure to do so will incur a supplemental one-hour charge.

If no one is booked after your slot, imaging may continue. Please record this extra time as overtime in the manual logbook.

A reservation can be changed or cancelled any time, up to 24 hours before a booking.

To cancel, email noushin.ahmadpour@umanitoba.ca.

When cancelling, especially on short notice, it is your responsibility to ensure that a system was not left on for you by the previous scheduled user. Failure to do so will result in a charge equal to the duration the system was left on.

Reservations not cancelled will be charged for the time entered into the online calendar at the time of booking.