Microscopy

Zeiss ELYRA LSM 710 / Axio Observer

The Elyra combines three dimensional (PRILM Technology), dual channel dSTORM/PALM super-resolution imaging with standard confocal microscopy to study protein localization and geometry at the nanoscale level. The setup is able to achieve an experimental resolution of 30 nm laterally, and 70 nm axially. The microscope is also suitable for live imaging of fluorescent-tagged transgenic cell lines.

Super-resolution laser lines: 405-488-542-643 nm.
Confocal laser lines: 458-488-514-543-633 nm.


Top Left: An isolated cardiomyocyte labelled for ryanodine receptors (RyR) imaged in super-resolution (dSTORM) and in standard wide-field microscopy (Image: Xin Shen, Postdoc). Top Right: Dual Colour 2D dSTORM imaging of L-type Ca2+ Channel (red) and RyR (blue) reveals surprisingly low degree of colocalization of two proteins (Image: Ornella Manfra, Postdoc). Bottom: Correlative 3D dSTORM and confocal imaging enables visualization of dyadic junctions in cardiomyocytes. Combining dSTORM-derived ryanodine receptor positions (yellow) with confocal images of t-tubules (blue) allows appreciation of the 3D complexity and irregularity of dyadic arrangements (Image: Xin Shen, Postdoc, IEMR).

Zeiss LSM 800 Airyscan / Axio Observer

Inverted super-resolution confocal microscope used for studying protein localization, motility and colocalization using traditional antibody labelling. Additionally, the microscope is fitted with an incubation chamber for live-cell imaging experiments requiring control of temperature and O2/CO2 levels. The LSM 800 enables simultaneous imaging of up to 4 different proteins. With Airyscan processing, the setup is able to achieve a resolution of 120 nm laterally and 350 nm axially, far exceeds that of a conventional confocal microscope.

Laser lines: 405-488-561-640 nm.


Top: An isolated rat cardiomyocyte labelled with Di-8-ANEPPS illustrating its extensive transverse tubular (TT) network. Bottom left: Cultured HL-1 cells labelled for DAPI (blue), BIN-1 (green) and α-tubulin (red). BIN-1 is a known to be an important modulator of TT growth and maintenance. Bottom right: Culture HL-1 cells labelled for Myotubularin, MTM1 (red) and microtubule (green). The absence of MTM-1 has been postulated to play a role in microtubule-mediated TT pathologies. (Images: Michael Frisk, Postdoc, IEMR).

Zeiss 7 Live / Axio Observer

Inverted confocal microscope used for studying Ca2+ dynamics in live cells. The 7 Live provides high speed scanning and gives acquisition rates of 1000 images of 512 x 512 pixels in less than 10 seconds. The microscope is set up with a perfusion system (37°C), and field stimulation. Flash photolysis equipment is also available.

Laser lines: 488-532 nm.


Top: An isolated rat cardiomyocyte labelled with the voltage sensitive dye FluoVolt. Fluorescence intensity sharply changes as the cell responds to electrical field stimulation (Image: David Lipsett, PhD Candidate). Bottom: An isolated rat cardiomyocyte labelled with the Ca2+ indicator Fluo-4. At rest, a hyperphosphorylated cell exhibits spontaneous local releases of Ca2+, giving rise to what is known as Ca2+ sparks. The summation of these microscopic events often leads to the generation of proarrhythmic Ca2+ wave (Image: Michael Frist, postdoc, IEMR).

Zeiss Observer D1 / Cell Tester

This fluorescence system enables wide-field fluorescence detection such as Fluo-4 (Easy Ratio Pro) by high-speed camera (Orca Flash 4.0, Hamamatsu Photonics) with WarpDrive interface controller. The setup is also equipped with a Cell Tester system (World Precision Instruments), enabling concomitant measurements of local cellular Ca2+ dynamics as well as contractile force development at varying sarcomere lengths. Additionally, the setup contains a heated perfusion system, as well as equipment for electrophysiology (Axon Axoclamp 200B) and field stimulation.


Top: An isolated rat cardiomyocyte attached to two glass fibers coated with MyoTak (a biocompatible cellular adhesive). Bottom: Developed force can be measured as the cell is stretched to different lengths. (Images: Jia Li, IEMR).

Zeiss LSM 510 / Axiovert 100

This inverted point-scanning confocal microscope has modest spatial and temporal resolution, and is commonly employed for both imaging of ionic fluorophores and static imaging protein localization / colocalization. This setup is also equipped with whole-cell fluorescence with PMT-based detection (Ratio Master, Photon Technology International), a heated perfusion system, electrophysiology (Axon Axoclamp 2B) and

Laser lines: 488-543-633 nm.




Calcium transients recorded using confocal line-scan from control and cultured isolated cardiomyocytes. Transverse tubule density progressively reduces during culture. This loss eventually results in dyssynchronous Ca2+ release from the sarcoplasmic reticulum store (Image: William Louch, IEMR).

Zeiss Axiovert 200 / Cairn Optoscan

This inverted wide-field microscope is employed for studying ionic dynamics in live cells. Whole-cell fluorescence can be measured using a fluorescence system from Cairn Research (Optoscource / Optoscan, 1 detection channel). The setup is equipped with a heated perfusion system, as well as equipment for measuring cell shortening (Crescent electronics) and electrophysiology (Axon Axoclamp 2B) and for exciting cells by field stimulation.

Zeiss Axio Observer / PTI Wide-Field Fluorescence

This inverted wide-field microscope is employed for studying ionic dynamics in live cells. Whole-cell ratiometric fluorescence (for example Fluo-4, Fura-2, SBFI) can be measured using the Ratio Master system from Photon Technology International employing Felix software. The setup is equipped with a heated perfusion system, equipment for measuring cell shortening (Crescent electronics) and electrophysiology (Axon Axoclamp 2A) and for exciting cells by field stimulation.

Huygens deconvolution software from Scientific Volume Imaging

The core facility is licensed for use of deconvolution software from SVI which improves image quality by reversing optical distortion in the image.

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