{"id":250,"date":"2020-03-06T20:20:34","date_gmt":"2020-03-06T20:20:34","guid":{"rendered":"https:\/\/lab.research.sickkids.ca\/imagingfacility\/?page_id=250"},"modified":"2025-04-11T17:30:00","modified_gmt":"2025-04-11T17:30:00","slug":"microscopes","status":"publish","type":"page","link":"https:\/\/lab.research.sickkids.ca\/imagingfacility\/equipment\/microscopes\/","title":{"rendered":"Microscopes"},"content":{"rendered":"<div class=\"wpb-content-wrapper\"><p>[vc_row][vc_column][vc_row_inner equal_height=&#8221;yes&#8221; css=&#8221;.vc_custom_1456148051953{margin-top: 0px !important;}&#8221;][vc_column_inner css=&#8221;.vc_custom_1456148028687{padding-right: 35px !important;padding-left: 35px !important;background-color: #dd4949 !important;}&#8221;][vc_custom_heading text=&#8221;SUPER RESOLUTION&#8221; font_container=&#8221;tag:h1|text_align:left|color:%23ffffff&#8221; use_theme_fonts=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;MINFLUX&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]<\/p>\n<div class=\"page\" title=\"Page 30\">\n<div class=\"section\">\n<div class=\"layoutArea\">\n<div class=\"column\">\n<p>MINFLUX is a method of imaging that is able to synergistically combine the strengths of STED and PALM\/STORM to achieve an unprecedented 3D localization precision of 1-3 nm, and can also be applied to the recording of molecular trajectories with frequencies of up to 10 kHz. In MINFLUX imaging, fluorophores are individually switched in a manner similar to SMLM, allowing for the separation of overlapping signal at a single-molecule level. In turn, the fluorophore localization is accomplished using a movable structured excitation beam featuring a central intensity minimum (zero). Localization is performed by actively targeting the zero of the excitation donut. As the intensity minimum of the excitation beam approaches the fluorophore, there is a corresponding decrease in the emitted fluorescence per unit excitation power, thereby indirectly revealing the residual distance of the molecule to the excitation zero. Through a series of calculated iterations, the excitation minimum is brought as close as possible to the molecule until the detected fluorescence rate matches that of the background noise. At this point, the perfectly controlled position of the zero-excitation intensity becomes a proxy for the position of the emitter.<\/p>\n<\/div>\n<\/div>\n<\/div>\n<\/div>\n<p>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;473&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System name:<\/strong> Abberior MINFLUX<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>100x\/1.45 (Minflux, confocal)<\/li>\n<li>40x\/0.95 (Widefield, eyepiece)<\/li>\n<li>20x\/0.8 (Widefield, eyepiece)<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>5x APD:\u00a0 DAPI (422nm &#8211; 467nm); GFP (500nm &#8211; 550nm); Cy3 (580nm &#8211; 630nm); Cy5 Near (650nm &#8211; 685nm); Cy5 Far (685nm &#8211; 720nm); PMT (Alignment)<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>Minflux &#8211; 640 nm<\/li>\n<li>Confocal &#8211; 561 nm, 488 nm<\/li>\n<li>Activation &#8211; 405 nm<\/li>\n<\/ul>\n<p><strong>Acquisition Modes<\/strong><\/p>\n<ul>\n<li>2D<\/li>\n<li>3D<\/li>\n<li>Fast 2D tracking<\/li>\n<li>3D tracking<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Structured Illumination Microscopy (SIM)&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]<\/p>\n<p class=\"Body\"><span lang=\"EN-US\">SIM is an imaging technique that employs patterned excitation light to generate interference patterns (moir\u00e9 patterns) that contain high frequency spatial information. By collecting a series of images with different interference patterns, specialized algorithms are able to extract super-resolution detail to produce a reconstructed final image that has approximately twice the resolution of traditional diffraction limited microscopes, ie. approx. 110 nm (XY), 300 nm (Z). This technique is ideally suited for 3D imaging of thin (&lt;20 um), fixed samples that have excellent fluorescence labeling.<\/span><\/p>\n<p>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;268&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System name:<\/strong> Zeiss Elyra PS1<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>63x\/1.4 PlanApo<\/li>\n<li>100x\/1.46 alpha PlanApo<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>Andor iXon3 885 (SIM)<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm (150 mW)<\/li>\n<li>488 nm (200 mW)<\/li>\n<li>561 nm (200 mW)<\/li>\n<li>640 nm (150 mW)<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Localization Microscopy (STORM\/PALM)&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]Stochastic optical reconstruction microscopy (STORM) and photoactivated localization microscopy (PALM) function by stochastically cycling fluorophores between \u2018on\u2019 and \u2018off\u2019 states in a controlled manner, followed by precise localization of the centre of each fluorescent molecule to within tens of nanometers. By processing thousands of images, the molecular coordinates of all labeled molecules are obtained with a resolution ten times better than traditional diffraction limited microscopes, i.e. approximately 20 nm (XY), 50 nm (Z). This technique is best suited for imaging the stoichiometric architecture of proteins within defined complexes, and is largely limited to fixed, cellular samples.[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;268&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Zeiss Elyra PS1<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>63x\/1.4 PlanApo<\/li>\n<li>100x\/1.46 alpha PlanApo<\/li>\n<\/ul>\n<p><strong>Cameras<\/strong><\/p>\n<ul>\n<li>Andor iXon DU897 (PALM)<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm (150 mW)<\/li>\n<li>488 nm (200 mW)<\/li>\n<li>561 nm (200 mW)<\/li>\n<li>640 nm (150 mW)<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>3D: double-helix PSF<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Stimulated Emission Depletion (STED)&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]<\/p>\n<p class=\"Body\"><span lang=\"EN-US\">STED microscopy overcomes the diffraction limit by overlapping the primary excitation laser with a secondary, donut-shaped, high-power laser beam. Irradiation by the secondary laser forces molecules in the outer region of the primary excitation area to return to the ground state, while leaving fluorophores in the centre of the excitation focus to form the high resolution image. This application can achieve resolutions up to 50 nm laterally, and up to 120 nm axially, and is best suited for 2D or 3D imaging of thin samples (&lt;50 um) that have excellent fluorescence labeling.<\/span><\/p>\n<p>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;467&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Leica Stellaris 8 FALCON w\/STED<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>93x\/1.3 STED (motCORR)<\/li>\n<li>100x\/1.4 STED<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>white light laser (pulsed, 440-790 nm)<\/li>\n<li>775 nm (pulsed, STED)<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>TauSTED<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][\/vc_row][vc_row anchor=&#8221;#confocal&#8221;][vc_column][vc_row_inner equal_height=&#8221;yes&#8221; css=&#8221;.vc_custom_1456148051953{margin-top: 0px !important;}&#8221;][vc_column_inner css=&#8221;.vc_custom_1456148028687{padding-right: 35px !important;padding-left: 35px !important;background-color: #dd4949 !important;}&#8221;][vc_custom_heading text=&#8221;CONFOCAL&#8221; font_container=&#8221;tag:h1|text_align:left|color:%23ffffff&#8221; use_theme_fonts=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_column_text]<span lang=\"EN-US\">Confocal microscopy is a powerful technique in which a pinhole is used to block out-of-focus light in image formation, thereby generating improved contrast and resolution compared to widefield microscopes. Generally speaking, there are two types of confocal microscopes: <strong>point scanning<\/strong> and <strong>spinning disk<\/strong>. Although slower than a spinning disk confocal, point scanning confocal microscopes tend to achieve higher resolutions, and have more integrated solutions for specialized techniques such as fluorescence recovery after photobleaching (FRAP). On the other hand, spinning disk confocals tend to have significant advantages in both speed and phototoxicity, making them ideally suited for live cell imaging.<\/span><\/p>\n<p><span lang=\"EN-US\">In recent years, the resolution of traditional point scanning confocal microscopes has been extended to the super-resolution realm by implementing three key components to a workflow: <\/span><\/p>\n<ol>\n<li><span lang=\"EN-US\">Closing the pinhole diameter below 1 Airy Unit, <\/span><\/li>\n<li><span lang=\"EN-US\">Using low-noise, high-sensitivity detectors,<\/span><\/li>\n<li><span lang=\"EN-US\">Combining pixel oversampling with modern deconvolution algorithms. <\/span><\/li>\n<\/ol>\n<p><span lang=\"EN-US\">Taken together, these implementations can improve resolution by up to 40% compared to traditional confocal microscopes, i.e. 120 nm (XY), 400 nm (Z), and can be applied to most standard confocal applications.<\/span><\/p>\n<p><span lang=\"EN-US\">A specialized application of confocal microscopy is <strong>multiphoton imaging<\/strong>. In this technique, a specialized infrared laser is used to selectively excite molecules located within a femtoliter volume of space, up to one millimetre deep within a sample. The primary advantage of multi photon imaging is reduced phytotoxicity and deep sample penetration, making it ideal for intravital microscopy.<\/span>[\/vc_column_text][vc_custom_heading text=&#8221;Point scanning confocals&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;281&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Leica SP8 Lightning Confocal\/Light Sheet<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>5x\/0.15<\/li>\n<li>20x\/0.75<\/li>\n<li>40x\/1.3 (motCORR)<\/li>\n<li>63x\/1.3<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>448 nm<\/li>\n<li>488 nm<\/li>\n<li>552 nm<\/li>\n<li>638 nm<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>4x HyD, 1x PMT, resonance scanner<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;474&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System name:<\/strong> Leica Stellaris 5 WLL<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>5x\/0.15<\/li>\n<li>10x\/0.4<\/li>\n<li>20x\/0.75<\/li>\n<li>40x\/1.1<\/li>\n<li>63x\/1.4<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>white light laser (pulsed 485-685 nm)<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>4x HyD S<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>Resonance scanner (2048 x 2048)<\/li>\n<li>Lightning Deconvolution<\/li>\n<li>TauContrast<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;467&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System name:<\/strong> Leica Stellaris 8 FALCON w\/STED<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>10x\/0.4<\/li>\n<li>20x\/0.75<\/li>\n<li>40x\/1.3<\/li>\n<li>63x\/1.4<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>white light laser (pulsed 440-790 nm)<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>2x HyD S, 2x HyD X, HyD R<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>Lightning Deconvolution<\/li>\n<li>TauContrast<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;294&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Zeiss LSM880 Airyscan<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>5x\/0.16<\/li>\n<li>10x\/0.45<\/li>\n<li>20x\/0.8 PlanApo<\/li>\n<li>40x\/1.1<\/li>\n<li>63x\/1.4 PlanApo<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>argon (458\/476\/488\/496\/514 nm)<\/li>\n<li>552 nm<\/li>\n<li>642 nm<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>\u00a032-channel spectral PMT, Airyscan Fast<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;275&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Nikon A1R Confocal Microscope w\/FCS + FLIM<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>10x\/0.5<\/li>\n<li>20x\/0.75<\/li>\n<li>40x\/1.25<\/li>\n<li>60x\/1.4<\/li>\n<li>100x\/1.4<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>402 nm<\/li>\n<li>440 nm (pulsed)<\/li>\n<li>488 nm<\/li>\n<li>561 nm<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>2 x GaAsP PMT, 2 x PMT<\/li>\n<li>32-channel spectral PMT<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>HD resonance scanner (1024 x 1024)<\/li>\n<li>Denoise.ai, NIS-Elements Deconvolution<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Spinning disk confocals&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;270&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Quorum Spinning Disk 2<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>10x\/0.4<\/li>\n<li>20x\/0.75<\/li>\n<li>40x\/0.9<\/li>\n<li>60x\/1.27<\/li>\n<li>60x\/1.35<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>491 nm<\/li>\n<li>561 nm<\/li>\n<li>642 nm<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>Hamamatsu C9100-13 EM-CCD<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;382&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name: <\/strong>Quorum Spinning Disk 3<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>10x\/0.4<\/li>\n<li>20x\/0.8<\/li>\n<li>40x\/1.1<\/li>\n<li>40x\/1.3<\/li>\n<li>63x\/1.4<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>491 nm<\/li>\n<li>561 nm<\/li>\n<li>637 nm<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>Hamamatsu C9100-13 EM-CCD, Photometrics Prime 95B<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;267&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Quorum Spinning Disk 4<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>4x\/0.16<\/li>\n<li>10x\/0.4<\/li>\n<li>20x\/0.75<\/li>\n<li>40x\/0.95<\/li>\n<li>60x\/1.35<\/li>\n<li>100x\/1.4<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>491 nm<\/li>\n<li>561 nm<\/li>\n<li>642 nm<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>Hamamatsu C9100-13 EM-CCD<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Multiphoton imaging&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;279&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Leica SP8 Confocal w\/Two-Photon<\/p>\n<p><strong>Objective<\/strong><\/p>\n<ul>\n<li>25x\/0.95<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>1 x HyD, 2 x PMT, 2 x NDD-HyD<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>Argon (458\/476\/488\/496\/515 nm)<\/li>\n<li>561 nm<\/li>\n<li>633 nm<\/li>\n<li>Coherent Ultra II (680-1080 nm, &gt;3.3 W)<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][\/vc_row][vc_row anchor=&#8221;#widefield&#8221;][vc_column][vc_row_inner equal_height=&#8221;yes&#8221; css=&#8221;.vc_custom_1456148051953{margin-top: 0px !important;}&#8221;][vc_column_inner css=&#8221;.vc_custom_1456148028687{padding-right: 35px !important;padding-left: 35px !important;background-color: #dd4949 !important;}&#8221;][vc_custom_heading text=&#8221;WIDEFIELD&#8221; font_container=&#8221;tag:h1|text_align:left|color:%23ffffff&#8221; use_theme_fonts=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_column_text]<span lang=\"EN-US\">Widefield imaging is the method of choice for basic fluorescence imaging. It is most often applied to examining protein expression levels in cells and tissue, general patterns of localization, and population studies. It is especially powerful for high-speed imaging, as well as ratiometric measurements of intracellular pH, or other ion levels. Finally, it is the tool of choice for researchers looking to image cellular samples with white light (eg. DIC imaging of wound assays, beating cilia, etc.).<\/span>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;452&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Nikon Epifluorescence Microscope w\/TIRF<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>4x\/0.2<\/li>\n<li>10x\/0.45<\/li>\n<li>20x\/0.8<\/li>\n<li>40x\/1.25 (Sil)<\/li>\n<li>40x\/1.3 Super Fluor (O)<\/li>\n<li>60x\/1.4 (O)<\/li>\n<li>60x\/1.49 (O) TIRF<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>Hamamatsu Orca Fusion BT (Fluorescence)<\/li>\n<li>Nikon DS10 (Colour)<\/li>\n<\/ul>\n<p><strong>Light Source<\/strong><\/p>\n<ul>\n<li>Spectra III (Excitation: 348 nm, 380 nm, 434 nm, 475 nm, 511 nm, 555 nm, 637 nm, 748 nm)<\/li>\n<li>Lasers (TIRF): 488 nm (90 mW), 561 nm (70 mW)<\/li>\n<\/ul>\n<p><strong>Filter Cubes<\/strong><\/p>\n<ul>\n<li>Fura2\/TRITC, Penta (390\/475\/555\/635\/747), Triple (440\/510\/575), TIRF (488\/561)<\/li>\n<li>Emission Filters: 432\/36, 474\/27, 515\/30, 544\/24, 595\/31, 680\/42, TIRF 525\/50, TIRF 600\/50<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;445&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Zeiss AxioZoom V16<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>1.0x\/0.25<\/li>\n<li>2.3x\/0.57<\/li>\n<\/ul>\n<p><strong>Zoom<\/strong><\/p>\n<ul>\n<li>0.7x &#8211; 11.2x<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>pco.edge 4.2 bi (Fluorescent)<\/li>\n<li>Axiocam 305 color (Colour)<\/li>\n<\/ul>\n<p><strong>Filter Cubes<br \/>\n<\/strong><\/p>\n<ul>\n<li>DAPI<\/li>\n<li>GFP<\/li>\n<li>RFP<\/li>\n<li>Cy5<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][\/vc_row][vc_row anchor=&#8221;#other&#8221;][vc_column][vc_row_inner equal_height=&#8221;yes&#8221; css=&#8221;.vc_custom_1456148051953{margin-top: 0px !important;}&#8221;][vc_column_inner css=&#8221;.vc_custom_1456148028687{padding-right: 35px !important;padding-left: 35px !important;background-color: #dd4949 !important;}&#8221;][vc_custom_heading text=&#8221;OTHER&#8221; font_container=&#8221;tag:h1|text_align:left|color:%23ffffff&#8221; use_theme_fonts=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Light sheet fluorescence microscopy&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]In light sheet imaging, samples are illuminated perpendicularly to the direction of observation using a thin sheet of light that is projected through the sample. Although the lateral resolution of a light sheet microscope is comparable to a widefield microscope, its key advantages are greatly reduced phototoxicity, rapid imaging, and the ability to image large samples from multiple directions for increased clarity and resolution. This technique has been masterfully adapted for use with cleared tissues, such as brain, kidney, and other organs measuring up to several millimetres in thickness[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;265&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Zeiss Light Sheet Z1<\/p>\n<p><strong>Illumination Objectives<\/strong><\/p>\n<ul>\n<li>5x\/0.1<\/li>\n<li>10x\/0.2<\/li>\n<\/ul>\n<p><strong>Detection Objectives<\/strong><\/p>\n<ul>\n<li>5x\/0.16<\/li>\n<li>10x\/0.5<\/li>\n<li>20x\/1.0<\/li>\n<li>20x\/1.0 (CLARITY, RI=1.45)<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>488 nm<\/li>\n<li>561 nm<\/li>\n<li>638 nm<\/li>\n<\/ul>\n<p><strong>Cameras<\/strong><\/p>\n<p>pCO Edge 5.5 x 2[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;449&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Miltenyi Biotec UM Blaze<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>1.1x\/0.1<\/li>\n<li>4.0x\/0.35<\/li>\n<li>12x\/0.53<\/li>\n<\/ul>\n<p><strong>Dipping Caps<\/strong><\/p>\n<ul>\n<li>1.1x DC40 (RI 1.33-1.49)<\/li>\n<li>1.1x DC57 (RI 1.50-1.57)<\/li>\n<li>4.0x DC33 (RI 1.33 &#8211; 1.41)<\/li>\n<li>4.0x DC49 (RI 1.42-1.48)<\/li>\n<li>4.0x DC57 (RI 1.49-1.57)<\/li>\n<li>12x DC33 (RI 1.33-1.41)<\/li>\n<li>12x DC49 (RI 1.42-1.48)<\/li>\n<li>12x DC57 (RI 1.49-1.57)<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>488 nm<\/li>\n<li>561 nm<\/li>\n<li>639 nm<\/li>\n<li>785 nm<\/li>\n<\/ul>\n<p><strong>Cameras<\/strong><\/p>\n<p>pco.edge 4.2[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;450&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Zeiss Lattice Light Sheet Microscope<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>llumination Lens: 13.3x\/0.4 (30 degrees)<\/li>\n<li>Detection Lens: 44.8x\/1.0 (60 degrees)<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>pco.edge 4.2<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>488 nm<\/li>\n<li>561 nm<\/li>\n<li>640 nm<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;281&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Leica SP8 Lightning Confocal\/Light Sheet<\/p>\n<p><strong>Illumination Objectives<\/strong><\/p>\n<ul>\n<li>6x\/0.05<\/li>\n<li>5x\/0.07<\/li>\n<li>5x\/0.15<\/li>\n<\/ul>\n<p><strong>Detection Objectives<\/strong><\/p>\n<ul>\n<li>5x\/0.15<\/li>\n<li>10x\/0.3<\/li>\n<li>25x\/0.95<\/li>\n<\/ul>\n<p><strong>TwinFlect Mirrors<\/strong><\/p>\n<ul>\n<li>5 mm<\/li>\n<li>5 mm<\/li>\n<li>8 mm<\/li>\n<li>8 mm (glycerol)<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>405 nm<\/li>\n<li>448 nm<\/li>\n<li>488 nm<\/li>\n<li>552 nm<\/li>\n<li>638 nm<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>\u00a0pCO Edge 5<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Fluorescence Lifetime Imaging (FLIM)&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]FLIM is an excellent technique for mapping the local environment and molecular interactions of a fluorophore. A fluorophore\u2019s lifetime (ie. the amount of time spent in the excited state) is a quantitative signature that can be used to extract information about its surrounding environment: ion concentrations, pH, viscosity, and molecular interactions. Furthermore, because fluorescence lifetime measurements are not affected by probe concentrations or photobleaching, and are independent of excitation, FLIM is an ideal tool for measuring FRET in living cells.[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;467&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:\u00a0<\/strong>Leica Stellaris 8 FALCON w\/STED<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>20x\/0.8<\/li>\n<li>40x\/1.3<\/li>\n<li>60x\/1.4<\/li>\n<li>93x\/1.3<\/li>\n<li>100x\/1.4<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>white light laser (pulsed, 440-790 nm)<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>2x HyD S, 2x HyD X, HyD R<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>Leica FALCON FLIM module<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Fluorescence Correlation Spectroscopy (FCS)&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]<span lang=\"EN-US\">FCS is a correlative analysis of fluorescence fluctuations over time within a defined volume. As fluorophores enter and exit the observed volume, the resulting fluctuations in intensity encode parameters such as diffusion rates, molecular volume, binding affinities, and colocalization. This tool is especially useful for measuring the mobility of rapidly diffusing cytosolic proteins.<\/span>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;467&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:\u00a0<\/strong>Leica Stellaris 8 FALCON w\/STED<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>40x\/1.1<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>white light laser (pulsed, 440-790 nm)<\/li>\n<\/ul>\n<p><strong>Detectors<\/strong><\/p>\n<ul>\n<li>2x HyD S, 2x HyD X, HyD R<\/li>\n<\/ul>\n<p><strong>Other<\/strong><\/p>\n<ul>\n<li>Leica FALCON FCS module<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Total internal reflection fluorescence (TIRF)&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]<span lang=\"EN-US\">TIRF microscopy is a specialized technique that allows for imaging of fluorescent molecules located within 200 nm of the coverslip. It accomplishes this by the generation of a fluorescent evanescent wave that decays exponentially as it projects upward from the surface of the coverslip. In doing so, only fluorophores located 100-200 nm from the coverslip are excited, resulting in very high signal-to-noise imaging of the basolateral surface of cells. This technique is especially useful for observing membrane associated processes such as ligand binding, cell adhesion, and vesicle trafficking from the cell surface.<\/span>[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;453&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> Nikon Epifluorescence Microscope w\/ TIRF<\/p>\n<p><strong>Objective<\/strong><\/p>\n<ul>\n<li>60x\/1.49 (O)<\/li>\n<\/ul>\n<p><strong>Camera<\/strong><\/p>\n<ul>\n<li>Hamamatsu ORCA-Fusion BT<\/li>\n<\/ul>\n<p><strong>Lasers<\/strong><\/p>\n<ul>\n<li>491 nm<\/li>\n<li>561 nm<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;Automated slide scanning&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text]Digital slide scanning is the ideal solution for imaging whole tissue sections. Up to 250 slides can be automatically scanned in a high-throughput fashion using high-resolution 20x and 40x objectives, in both brightfield and fluorescence modes. The resulting images can be analyzed to provide area quantification, complex colour analysis, cytonuclear quantitation, etc. This technology is featured as a service in the Facility, with the turnaround typically being 24 hours.[\/vc_column_text][vc_row_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_single_image image=&#8221;339&#8243; img_size=&#8221;large&#8221;][\/vc_column_inner][vc_column_inner width=&#8221;1\/2&#8243;][vc_column_text]<strong>System Name:<\/strong> 3DHistech Pannoramic Flash II Slide Scanner<\/p>\n<p><strong>Objectives<\/strong><\/p>\n<ul>\n<li>20x\/0.8<\/li>\n<li>40x\/0.95<\/li>\n<\/ul>\n<p><strong>Cameras<\/strong><\/p>\n<ul>\n<li>pCO Edge 4.2 sCMOS<\/li>\n<li>CIS VCC-FC60FR19CL<\/li>\n<\/ul>\n<p><strong>Fluororescence Filter Cubes<\/strong><\/p>\n<ul>\n<li>DAPI<\/li>\n<li>GFP<\/li>\n<li>Cy3<\/li>\n<li>Cy5<\/li>\n<li>Quad cube<\/li>\n<\/ul>\n<p>[\/vc_column_text][\/vc_column_inner][\/vc_row_inner][\/vc_column][vc_column][vc_column_text]<strong>All Imaging Facility equipment can be booked through <a href=\"https:\/\/my.qreserve.com\/site\/Y0_vzAyNEIg0VvHbjraXvEixxYg=\" target=\"_blank\" rel=\"noopener noreferrer\">QReserve<\/a>.<\/strong>[\/vc_column_text][\/vc_column][\/vc_row]<\/p>\n<\/div>","protected":false},"excerpt":{"rendered":"<p>[vc_row][vc_column][vc_row_inner equal_height=&#8221;yes&#8221; css=&#8221;.vc_custom_1456148051953{margin-top: 0px !important;}&#8221;][vc_column_inner css=&#8221;.vc_custom_1456148028687{padding-right: 35px !important;padding-left: 35px !important;background-color: #dd4949 !important;}&#8221;][vc_custom_heading text=&#8221;SUPER RESOLUTION&#8221; font_container=&#8221;tag:h1|text_align:left|color:%23ffffff&#8221; use_theme_fonts=&#8221;yes&#8221;][\/vc_column_inner][\/vc_row_inner][vc_custom_heading text=&#8221;MINFLUX&#8221; font_container=&#8221;tag:h3|text_align:left&#8221; use_theme_fonts=&#8221;yes&#8221;][vc_column_text] MINFLUX is a method of imaging that is able to synergistically combine the strengths of STED and PALM\/STORM to achieve an unprecedented 3D localization precision of 1-3 nm, and can also be applied to the recording&hellip;<\/p>\n","protected":false},"author":267,"featured_media":0,"parent":27,"menu_order":0,"comment_status":"closed","ping_status":"closed","template":"","meta":{"footnotes":""},"class_list":["post-250","page","type-page","status-publish","hentry","description-off"],"yoast_head":"<!-- This site is optimized with the Yoast SEO Premium plugin v27.0 (Yoast SEO v27.0) - https:\/\/yoast.com\/product\/yoast-seo-premium-wordpress\/ -->\n<title>Microscopes - SickKids Imaging Facility<\/title>\n<meta name=\"robots\" content=\"index, follow, max-snippet:-1, max-image-preview:large, max-video-preview:-1\" \/>\n<link rel=\"canonical\" href=\"https:\/\/lab.research.sickkids.ca\/imagingfacility\/equipment\/microscopes\/\" \/>\n<meta property=\"og:locale\" content=\"en_US\" \/>\n<meta property=\"og:type\" content=\"article\" \/>\n<meta property=\"og:title\" content=\"Microscopes\" \/>\n<meta property=\"og:description\" content=\"[vc_row][vc_column][vc_row_inner equal_height=&#8221;yes&#8221; 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