Normally LAS will be running and HyVolution2 mode already selected. Leica DMi8 - University of Rochester Medical Center Leica LAS NAVIGATOR -- a wizard for both tile scan & stitching and for multi-well plates -- can be used without selecting any of these 'templates'. This laser based confocal system permits measurements of fluorescently labeled tissue sections, individual cells, or particles adhered to matrices over a wide selection of visible wavelengths. Data range: we normally operate the SP8 in 16-bit mode (HyVolution configuration setting), which results in dynamic range of0 .. 65535 (which is 2^16 - 1, since zero is the first value). In addition, the Leica TCS SP8 features 3 HyDs (hybrid detectors combining the characteristics of a classic PMT with a highly sensitive avalanche photodiode) and 1 PMT which can measure real time response of time-critical sequences due to its fast scan speed. Probably more important to refractive index match your choice of mounting medium (ex. For example, 1800 Hz requires zoom 7.5x or more (you control number of pixels). The Facility features a 2019 Leica SP8 Confocal Microscope with White Light Laser and Fluorescence Lifetime Imaging Microscopy (FLIM) capability. 100 to 300 300 needs to use 16-bit mode [12-bit would work, but the image file is going to be 16-bit inside LAS X and export to raw TIFF, so might as well use 16-bit format). Cambridge, MA 02139-4307 When done with NAVIGATOR, please return to HyVolution2, turn off "red chain lock", and set speed to 600 Hz (from 700 Hz). All lasers to stand by For standard applications, use the easy to install filter cubes with automatic RFID identification. Both 5 and 8 are fast FLIM (fluorescence lifetime imaging microscopy) capable, 5 with TauSense modes (TauSeparation looks most useful to me; I note "5" does not provide raw data dump, so biophysicists may be bummed out if only have access to "5", but most biologists do not need the massive amount of raw data deluge of "8', and the money could be spent on more detectors for the "5"), 8 with "full data deluge" (~125 time intervals of ~100 picoseconds [0.1 ns] per 12.5 nanosecond pulse interval), TauSense and also FALCON (fluorescence lifetime contrast) and lifetime Phasor plots (set phasor to stun! When you change the contrast method, the microscope automatically adapts the illumination settings, parfocality, brightness, and diaphragm position to that method. Lightning (adaptive deconvolution) Summary of Instrument Features: Microscope: Inverted Leica DMi8 Microscope Software: LAX Software Objectives: Standard: 93x/1.30 - Glyc - HC PL Apochromat C52 63x/1.40 - Oil - HC PL Apochromat C52 20x/0.75 . (ii) if non-ACCM users add up to more than 20 hours in any work week and ACCM users did not need 20 hours: great for everyone. If used 63x oil gently wipe oil off the 63x lens with lens paper as I taught you (never use kimwipes or paper towels on our lenses). You will find a more detailed list of local contacts here. Huygens Essential deconvolution (SVI Hyugens Essential intregrated into Leica LAS X software). or lab/dept administrator set them up with at least one cost object (account number) in Agilent iLab. Congratulations! Page 69: Brighteld (Tl) PI-TL" lter cube using LAS X software. Leica DMi8 with manual Transmitted Light method sic microscope settings. Leica SP8 inverted confocal microscope System description: DMi8, inverted microscope: BF, Fluo (blue, green, red) BF TL Detector (transmitted light image, no additional illumination or time) Scanning stage with z-Galvo and Navigator (overviews, selection of regions of interest,.) the model demo'd has some different capabilities than the two featured STELLARIS models (all 3 can have up to 5 S detectors or a ix of S and/or HyD 3rd generation visible GaAs or NIR :GaAsP detectors) (note: STELLARIS uses an S detector for field service engineer calibrations, so no needfor any PMT re: SP8). You can save in HDF5 (32-bit loating point values), which Fiji ImageJ can open. Like a GPS for your cells, the software moduleLAS X Navigatorensures that you always have a clear roadmap to brilliant data. Prolong Glass) and high NA lens immersion medium on this microscope, Leica oil R.I. 1.518. All our pre-owned Microscopes Confocal include a worry-free 180-day warranty. Z all the way down. For Widefield applications, call or email Paula Cranfill: No matter which of the instrument components are part of the experiment, the system will run at the highest speed possible. I do not usually emphasize this value, since the user is controlling scan speed and image dimension, and this is the product (also, sometimes LAS X does not show the correct value!). Change from the macro (35 mm) to nano (200 nm) with only a click. Every DMi8 microscope can be equipped with up to two Infinity Ports, allowing direct access to the infinity space for flexible upgrading of your DMi8. McNamara 20220106H 20210302U ACCM Leica SP8 Confocal Microscope Quick Tips on Starting Finishing User Session. The DMi8 S is a versatile and userfriendly system that empowers biomedical researchers to probe molecular machineries within the cell with super-resolution, photo-manipulation and optogenetics. For more information,visithttp://www2.leica-microsystems.com/e/149501/jhu-lasx-/3tsp5w/258951420, For Confocal applications, call or email Geoff Daniels (senior sales): If you or anyone you know have a bunch of money to donate for us to buy a STELLARIS, that would be awesome (we would trade in the SP8 and uses the same space). CMN Core has a new Leica Stellaris 8 Confocal microscope. Price from $9.99to $1999.99 inverted leica dmi8 sp8 confocal microscope- by Bioz Stars, 2023-05 96/100stars Training courses: The graduate training course for the SP8 is BSC 411 Confocal Microscopy in Biology. Leica STED and confocal | Light Microscopy Core Facility Refractive index of the Leica immersion oil is 1.518. Most research lab PCs struggle to deal with >1 Gigabyte (GB) data files, so I recommend keeping each LIF container file to under 1 GB ex: save and close project file after each slide, potentially after each coverglass. . Leica SP8 confocal microscope | Ross Fluorescence Imaging Center And because the objectives are positioned below the stage the risk of collision with the sample is reduced. June 13, 2018:Leica SP8 is now in Ross S910A(S = Service corridor). If you get oil on the 20x lens, or it looks hazy compared to the 20x lens (i.e. Excellent objective lens. Confocal 1.0 Airy Unit at 500nm fluorescence emission: dxy = 0.51 * Lambda / NA = 0.51 * 500 nm / 1.3 = 182 nm. Leica DMi8 A System, Sample Preparation for Electron Microscopy. Filter cubes: DAPI, FITC, RHOD, FURA2, and a QUAD-S. We have a filter wheel for rapid changes in excitation and emission. Leica DMi8 inverted laser scanning confocal microscope with both galvonomic and resonance scanners and 5 spectral internal HyD-S detectors. McNamara et al 2014 MMB Low magnification confocal microscopy https://downloads.leica-microsystems.com/Leica%20TCS%20SP8/Brochures/Leica%20TCS%20SP8%20Scan%20Head-Flyer_EN.pdf, https://images.proteinatlas.org/35/674_H5_6_blue_red_green.jpg, Relative XY resolution improvement (approximate - Zeiss has a nice graph on this in a PDF brochure). "Rab7 and . I do note the HyD detectors have some background counts even with no laser power this is low and random (i.e. Confocal Microscopes | Medical School Durham, NC 27708lmcf@duke.edu There is an channel unmixing "matrix" to let you unmix 'spectral overlaps'. * $70/hr for users, with Dr. McNamara's oversight. 10 positions per second with a 40x objective. Example, 500-540 nm is away from 488 and 552 nm laser lines. Leica DMi8 M System, Do you want help to ensure reliable measurements? I estimate 10x advantage for each 3rd gen HyD at less than 2x the cost of each2nd gen HyD. The high speed linear motor stage offers absolutely precise positioning at previously unattainable speeds, e.g. An inverted laser scanning microscope for high resolution fluorescent imaging of fixed and live samples. Combined with an increased focus travel range of 12 mm, choose closed loop focus for the highest repeatability with multi-point time lapse experiments. The Infinity Port Connector, along with complete optomechanical design documentation, opens the Leica DMi8 light path to any accessory you want to add. See more of your sample in a shorter time thanks to the Macro* objective offered exclusively by Leica Microsystems. (iii) We encourage ACCM users to reserve well in advance (i.e.1+ week). Long term, the SP8 scanhead does not have an X1 port -- this is upgradable in the field. Contact us to find out how to upgrade your DMi8 with the latest DMi8 S platform options. 10); better image quality with more photons (ex. Koch Institute for Integrative Cancer Research You can evaluate each by enabling sequential scan tracks with each pinhole setting. Laser power more power gives more photons, but faster photobleaching and/or eventually reach either the maximum counting rate of the detector or the maximum photon flux of the fluorophores. Location TBD Contact Dane Jensen ( ddj3@nyu.edu) Imagine a world where everyone smiles Below: SVI.nl recommendations for the 63x/1.4NA lens, for Brilliant Violet 421: 405nm excitation, 440 nm emission. Staten Island, NY 10314 This new detector is a new design (ask your local Leica rep what is it), with the HyD(TM) now being just a trademark. Once you know what you are doing, you could acquire "over lunch" (or over long lab meeting), or overnight (or over weekend), unattended -- and use your time better that confocal sitting if you do not need to be one hand. Hardware zoom 0.75x - 48x (by changing confocal scanning area with the XY scanning mirrors). This novel design facilitates the integration of additional fluorescence light sources and laser systems for advanced applications like. Instructions for Use Gebrauchsanweisung Mode d'emploi, Leica Microsystems CMS GmbH Instructions 11934056, Revision 1.1 from 2015-07-15, Inverted research microscope for material testing (73 pages), Stand for neuro surgical microscopes (264 pages), Manual will be automatically added to "My Manuals", Safety Switch (Interlock) on the Microscope, Safety Hazard in the Condenser Area During Operation, Safety Hazard Due to Openings at Microscope, Note Regarding the Safety Hazards in Sections 3.3.3.1 and Section 3.3.3.2, Leica STP4000/8000 External Control Panel, Adjusting the Step Sizes on Motorized Stages, Differential Interference Contrast - DIC (TL), Integrated Modulation Contrast - IMC (TL), Leica Dmi8 Congurations Instrument Overview, Microscope Leica DMi8 A Installation Manual, Microscope Leica DMI4000B Instructions Manual, Microscope Leica DMI3000B Instructions Manual, Microscope Leica DMI6000B Operating Manual, Microscope Leica DM IRB Instructions Manual, Microscope Leica DMI6000 B Operating Manual, Microscope Leica DMi1 Instructions For Use Manual, Microscope Leica DMIL Instructions Manual, Microscope Leica DM IRM Instructions Manual, Microscope Leica DMI Series Instructions Manual, Page 6: Important Notes About This Manual, Page 8: Intended Purpose Of The Microscope, Page 15: Safety Hazard Due To Folded Back, Page 16: Safety Hazard Above The Condenser, Page 17: Safety Hazard Due To Openings At Microscope, Page 18: Note Regarding The Safety Hazards In Sections 3.3.3.1 And Section 3.3.3.2, Page 22: Notes On Handling The Touch Screen, Page 23: Leica Dmi8 Congurations Instrument Overview, Page 38: The Leica Smartmove Control Element, Page 39: Leica Stp4000/8000 External Control Panel, Page 54: Adjusting The Step Sizes On Motorized Stages, Page 62: Aperture Diaphragm And Field Diaphragm, Page 68: The Memory Function On The Touch Screen, Page 76: Differential Interference Contrast - Dic (Tl), Page 78: Integrated Phase Contrast - Iph (Tl), Page 80: Integrated Modulation Contrast - Imc (Tl), Page 93: Essential Consumable And Spare Parts, Page 97: Overview Of Touch Screen Pictograms. High-speed and high-sensitivity multi-modal confocal imaging with 2 pinhole sizes (25um and 40um) Environmental control for samples which need heating and CO2. The Hamamatsu Flash 4.0 sCMOS camera guarantees fast image acquisition with a large field of view (2048 x 2048 . Please note that Z-step size is the physical amount the Z-motor moves the objective lens. You will find a more detailed list of local contacts here. Total pixel dwell time (TPDT): product of line accumulation (ex. Tip: If you use immersion oil, clean off your slide with a dry kimwipe(s). One PMT (Hamamatsu R9624) with low dark current, 40 MHz sampling of ADC digitizer -- in practice is used by Leica field service engineer(s) for calibration during service visits (ex. Download the Leica software (LAS AF Lite-version 2.3.0) here. NYU.edu requires JavaScript be enabled in your browser in order to use important features of the site. This page was printed from http://flowcyt.rutgers.edu/instrumentation/leica-tcs-sp8-confocal-microscope/ at 6:14 PM Friday, June 2, 2023. 0.1 wt% fluorescein sodium salt was added to the PVA precursor as a fluorescent marker, and florescent HA-PVA hydrogels . Inverted Leica Dmi8 Sp8 Confocal Microscope - Bioz Note: 0.3 AU and "Area = pi r^2"implies 9% intensity. Undergraduates: BSC 356 is the undergrad SP8 training course, and runs concurrently with BSC 411; contact Dr. Edwards for an override. Strong tough hydrogels via the synergy of freeze-casting and salting On our SP8, you can sit with Dr. McNamara (and/or Leica representatives) and evaluate 'on image core time' the HyD's against each other (HyD1vs HyD3) and our standard PMT ("PMT2") with different settings (sequential track mode makes this easy). (i) If ACCM users get above 20 hours per work week: great. ), especially refractive index matching (1.518 for Leica 63x/1.4 NA objective lens, Leica immmersion oil (R.I. 1.518) and your mounting medium ThermoFisher Prolong Glass, after correct hardening process, comes very close you could also use "TDE" 2,2'-thiodiethanol, Staudt Hel 2006 MRT). It is a workhorse microscope that enables single molecule imaging all the way up to whole embryo imaging. We have the newer Klondike scanner withlinear scanning electronics, not the oldersinusoidal "Fermi" FOV scanner electronics (info on scanner electronics from Steffen Dietzel post on confocal listserv on 20220221Mon. Max image resolution 8192 x 8192 pixels (64 megapixels). Microscope:We acknowledge the Department of Anesthesiology & Critical Care Medicine (ACCM) for access to their Leica SP8 confocal microscope. Get the latest business insights from Dun & Bradstreet. That is, if imaging air (R.I. 1.0), physical step and "optical step" in the specimen are the same. 63x / 1.40 NA (oil immersion) pinhole 0.5 Airy Unit, 63x / 1.40 NA (oil immersion) pinhole 0.3 Airy Unit. Reserve theLeica SP8 Confocal MicroscopeRead theUser Manual, 2800 Victory Blvd Run your experiments up to 5x faster, integrate third party triggered components and synchronize the acquisition with the exposure to minimize photodamage. Go into Configuration, Settings, set data acquisition mode to 16-bit (not 8-bit), make sure HyD detectors are in Photon Counting mode, set line accumulation for each scan track to what you need (load causes each to go to 16; 10 is often enough, bright fluorophore labels or not-so-important channels, such as most users DAPI, can often be set to less, saving time and money). This tool of choice is suitable for demanding research applications and untrained operators alike. * I propose "Fast Photon Counting" (FPC) for now: 3rd gen HyDs, used with our current lasers, get data acquired a lot more quickly. Common 2*2 immunofluorescence experiment: scan track 1: 488 nm and 638 nm lasers, AF488 on HyD1, AF647 on HyD3. 1 transmitted light detector (brightfield, DIC). The scheduling will move to iLab when the ACCM's iLab page is activated. Follow all instructions! Second session GM walks user through loading settings, reminding safety features (make sure objective lens will not crash into specimen or holder). Confocal microscopy was carried out using a Leica DMi8 confocal microscope. Also possible to deconvolve in Huygens Essential. Leica Stellaris 8 w/ Leica DMI8 ID #CA-11959. We then pivot to the user specimen(s). It will be offered in Fall 2023. * We are very open to labs purchasing -- in exchange for time credit -- additional components for the Leica SP8 microscope. Talk to our experts. If you need super-resolution (beyonf standard of care confocal 1.0 or 0.66 Airy unit), consider using one of the other microscope facilities on campus -- i.e. Data is YOUR responsibility (and you are responsible to your P.I. For 20x/0.75NA objective lens: 1 Airy unit pinhole size, 120 nm XY pixel size, 360 nm Z-step size (GM's simple XY has three times better spatial resolution than Z). Combine Leica optical quality, a wide range of contrast modes, and intuitive software in one system to help you speed up your workflow. I note that Alexa Fluor PLUS secondary antibodies from ThermoFisher are 3x brighter, though 1.3x more expensive than plain AF antibodies. Leica SP8 Confocal Microscope | Microscopes, Imagers and Instruments Coronavirus (COVID-19): Latest Updates | Visitation PoliciesVisitation PoliciesVisitation PoliciesVisitation PoliciesVisitation Policies | COVID-19 Testing | Vaccine InformationVaccine InformationVaccine Information, Leica Stellaris 5 Inverted Confocal Microscope, 2023 University of Rochester Medical CenterRochester, NY, Clinical and Translational Sciences Institute, Monroe County Community Health Improvement Plan, Center for Advanced Light Microscopy and Nanoscopy, Zeiss Palm MicroBeam Laser Microdissection, Center for Advanced Light Microscopy and Nanoscopy (CALMN), Laser Scanning Confocal Microscopy (Standard and Resonant scanners available). If you think you have a better way, convince George you are right and we will change. June 1,2018: (see also above) Leica SP8 calendar/scheduler moved from Ross FIC (this site)to iLab Organizer as ofJune 1, 2018 (this scheduler booked all the time or calendar removed).***. The SP8 does enable sequential scan tracks, so you can image more than "3plex". Then set up high resolution image acquisition automatically using templates for slides, dishes and multiwell plates. https://help.ilab.agilent.com/36900-managing-your-group/279959-membership-requests-fund-numbers. Paula.Cranfill@leica-microsystems.com The Leica Application Suite (LAS) software lightens your workload by offering a variety of expert modules, e.g. Microscopy. Leica Application Suite X (LAS X) is the one software platform for all Leica microscopes: It integrates confocal, widefield, stereo, super-resolution, and light-sheet instruments from Leica Microsystems. We will continue to manage and host the microscope. Contact Us. Bioz Stars score: 86/100, based on 1 PubMed citations. We are happy to answer all your questions and concerns. Smaller pinhole sizes expected to reduce light level -- by how much is NOT simply proprotional to relative pinhole area: best to measure for yourself. A cool feature of STELLARIS confocal scan head is it can use FIVE (!!!!!) Leica DMi8 M / C / A Inverted Microscopes for Industry Invert The Game - And Stay Ahead Being ahead of the competition is what drives your business. Yes, you can acquire an ok image. * open access to JHU (and outside) researchers - if heavily booked, ACCM staff will have priority access (i.e. The lab section schedule will be determined once we know everyones availability. The "photon counting speedadvantage" of 3rd gen HyD's vs current 2nd gen is non-trivial to figure out under real use conditions. from 10x HDF to 50X DIC), Program one function key to acquire an image with the camera for documentation, Contrast techniques: Brightfield, Polorization, and Differential Interference Contrast (DIC), Manual 2-gear or 3-gear focus drive with focus stop and torque adjustment, 6-fold M32 coded or 6-fold M25 coded objective turret, 2-fold reflector turret coded, 6 fold reflector turret coded or uncoded, Contrast techniques: Brightfield, Darkfield, Polorization, Fluorescence, and Differential Interference Contrast (DIC), Motorized or 3-gear focus drive with focus stop and torque adjustment, 6-fold M32 motorized, 6-fold M32 coded, 6-fold M25 motorized or 6-fold M25 coded objective turret, 6-fold reflector turret motorized, 2-fold reflector turret coded, 6-fold reflector turret coded, 6-fold reflector turret uncoded, Exclusive manual or motorized UC-3D illumination, Contrasting techniques: Brightfield, Polorization, Fluorescence, and Differential Interference Contrast (DIC), Illumination and contrast management (diaphragm module). Each user should contact George McNamara (email gmcnamara@jhmi.edu) the fluorophores and specimens they plan to use. * FALCON = FAst Lifetime CONtrast (Fluorescence Lifetime on HyD photon counting detectors) ==> potential future upgrade of SP8). Leica SP8 scanhead schematic - We have LIAchroic (not AOBS) and two 2nd generation HyD detectors (we have rotation, we do not currently have X1 port). Andor Dragonfly Spinning Disk Confocal plus | Light Microscopy Core Massachusetts Institute of Technology this is even though a nice image can often be acquired with maximum (say) 100 photon counts; better would be 300; if you have over 1000 photon counts for bright pixels, you are probably spending more time acquiring than you need. If high image quality specimen, optimal image acquisition settings (at or near resolution limit), then deconvolution can improve resolution by ~10% and equally important, improve contrast and reduce haze. If our user base does not need all the "8's" capabilities, a "5" or a "as demo'd" model would still enable much more productivity than our workhorse Leica SP8 confocal microscope (that was purchased within budget constraints and sensible choice for functionality and limiting annual service contract cost). For instructions on how to enable JavaScript in your browser, NYU Dentistry Translational Research Center, NYU Dentistry Center for Oral Health Policy and Management, NYU Center for Skeletal and Craniofacial Biology (CSCB), NYU Dentistry Oral Health Center for People with Disabilities. 500 Main Street, Building 76 * Two excellent photon counting detectors, HyD1 and HyD3 (second generation hybrid detectors). Leica DMi8 (live) widefield fluorescence microscope for live cell imaging. Specifications. The DMi8 modular research microscope is the heart of the DMi8 S system. It is accessible from TCS SP8 mode only - the "many tiles" icon to the right of tile scan and mark&find ("NAV" is not visible in HyVolution2). ZERO BIAS - scores, article reviews, protocol conditions and more New user training notes (see also New User Training QA QC SOP doc): We normally train new users in two 2 hour sessions, ideally consecutive days. You can also look into DNA-PAINT and using our FISHscope for single molecule localization microscopy. These are (estimate) 40% Quantum Efficiency (QE) and were introduced Spring 2018. Fully configurable with manual to motorized components, the DMi8 microscope allows you to build the imaging system for your research and budget needs. Our 10x lens was not purchased for confocal microscopy. The Leica DMi8 is built with flexibility in mind. Deconvolution -- i.e. We would love to have 2, 4 or 8 APDs on the X1 port, to enable simultaneous 13plex (5 internal, 8 on X1) acquisition per laser line. ISU Leica SP8 Specifications - Confocal Microscopy Prof. Cyrus David Mintz, March 2018, 51 hours, in one time block). Save time and make sure your time lapse imaging isnt affected by changes in experimental conditions. LEICA DMI8 INSTRUCTIONS FOR USE MANUAL Pdf Download Do you need to work with big samples and analyse more of them? We hosted Leica STELLARIS confocal microscope demo with: * new "S" detectors (SiPM), ~55% QE (max) and good performance out to NIR. We typically recommend 10 line acumulation, resulting in 'typical' pixel (voxel) counts of 5 to 100 vs "empty" pixel dounts of zero or one or two. scan track 2:405 nm and 552nm lasers, DAPI on HyD1, AF568on HyD3. The DMi8 S is a flexible solution for advanced widefield research. A newer 64 bit version of LAS AF Lite (3.3.1) is available upon request. For example, 1 microsecond. Our SP8 has space for two more HyD's internally. More about LAS X Microscope Software for Life Sciences. Show local contacts, Leica DMi8 M / C / A Inverted Microscopes for Industry, Do you need a system for a simple and dedicated task? Microscopes > Confocal | Copia Scientific Used Lab Equipment MyJHU->Microsoft OneDrive or from our server immediately after your session). Only 91.84 degrees (the rotation angle that matches the eyepiece and XY stage) and 1.94 degrees (which "NAV" will default to) work correctly. Illinois State University Advanced Bioimaging Facility. MicFac -- that have found money for super-resolution. Leica "WLL" white light laser [super-continuum laser, 80 MHZ = 12.5 ns interval], to enable precised excitation wavelengths (ex. (919) 660.1234, Medical Sciences Research Building III Cluster, Andor XD Spinning Disk Confocal Microscope Manual, Andor Dragonfly Spinning Disk Confocal Microscope (User's Manual), DeltaVision Elite Deconvolution Microscope, Leica AM Total Internal Reflection Fluorescence Manual, Live Cell Station (Zeiss Axio Observer Z1) Manual - With MetaMorph, Live Cell Station (Zeiss Axio Observer Z1) Manual - Single or Tile Imaging With ZEN, Olympus VivaView FL Incubator Microscope Manual, ZEN 2.3 Pro: Single Field or Tiling/Stitching Multiple Fields, Zeiss Axio Imager Z2 Upright Microscope Manual, Zeiss MosaiX - Color brightfield tiling and image stitching on the LCM, Zeiss PALM MicroBeam Laser Capture Microdissection Manual, Paraformaldehyde, Formadehyde and Formalin, Stimulated Emission Depletion super-resolution imaging, High sensitivity point scanning confocal with pulsed white light laser, High-sensitivityGaAsPHyDdetectors with gating and photon counting, 3D z-stacks,timelapse, stitching, multi-positiontimelapse, STED depletion lasers: 592nm, 660nm, 775nm, Resonant scanner (8 000 Hz) - 28 fps at 512*512, Green (I3 = xBP470-490, dichroic 510, mLP515), Red (N2.1 = x515-560, dichroic 580, mLP590, 20x NA 0.75 HC PL APO multi-Imm Corr (oil, water, glycerol) DIC WD 270 um, 40x NA 1.3 oil HC PL APO CS2 OIL DIC WD 240 um, 93x NA 1.3 glycerol with motorized correction collar (for STED) WD 0.3mm, Five channels - two PMTs, three HyDs - high sensitivity Hybrid Detectors - PMT1 | HyD2 | PMT3 | HyD4 | HyD5, All spectral detectors; HyDs have gating for STED, Huygens deconvolution software linked to LAS X for STED deconvolution.
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