No matter which of the instrument components are part of the experiment, the system will run at the highest speed possible. 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. Leica DMi8 Confocal Microscope: Whether you need to precisely follow the development of a single cell in a dish, screen through multiple assays, obtain single molecule resolution, or tease out behaviors of complex processes, a DMi8 S system will enable you to see more, see faster, and find the hidden. Sometimes users do need big dynamic range! Specifications:Dimensions: (WxDxH) 335 x 100 x 175 cm (11 x 3.3 x 5.9 in. The instrument has extremely high sensitivity and can differentiate between closely emitting chromophores. See more of your sample in a shorter time thanks to the Macro* objective offered exclusively by Leica Microsystems.
[email protected] Our HyD's are "2nd generation", in mid-2018 Leica introduced single molecule detection SMD HyD's, that each count 2x faster than our 2nd gen HyD's. Combine Leica optical quality, a wide range of contrast modes, and intuitive software in one system to help you speed up your workflow. We will continue to manage and host the microscope. * Liachroic beamsplitters (similar to our SP8). For example, 37 C environmental unit (that works on SP8 / DMi8) and two more PMT detectors to "max out" the SP8 scan head. This would be an amazing microscope! Combination is rated for 140 nm XY resolution (acquire with 50nm or 40nm pixel size, typically Z = 3 * XY pixel size, so 150 or 120nm), compared to "conventional" confocal ~210 nm XY resolution (at 500 nm emission wavelength 405 nm excitation of any of BUV395, BV421, SB436 may enable better resolution). * open access to JHU (and outside) researchers - if heavily booked, ACCM staff will have priority access (i.e. Normally LAS will be running and HyVolution2 mode already selected. HyVolution2 quantitative deconvolution software using NVidia M6000 GPU card ("NEAR instant gratification" - some day we would like to upgrade to NVidia RTX 3080 or 3090 GPU, and new PCIe gen4 PC if you have lots of money, please cotnribute to our core). Get the latest business insights from Dun & Bradstreet. The instrument is equipped with 4 air cooled lasers allowing for excitation wavelengths of 405, 488, 552 and 638 nm. For Widefield applications, call or email Paula Cranfill: unusual specimens). or lab/dept administrator set them up with at least one cost object (account number) in Agilent iLab. Scanner with Scanfield Rotation 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). 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). Failure to sign in = Trespassing. GM can send PDF instructions and JHU iLab admin contacts if need help. I note that Alexa Fluor PLUS secondary antibodies from ThermoFisher are 3x brighter, though 1.3x more expensive than plain AF antibodies. Confocal sweetest spot: Tom Reece (NIDDK, NIH) on confocal listserv wrote that he often uses pinhole in range of 0.6 to 0.7 Airy unit, he calles "confocal sweet spot" (really confocal sweet range) to get better resolution (by a modest amount) and not much light loss (that ios, if one used infinitely small pinhole, would mathematically and geometrical optically get better resolution, but in practice, infinitely small number of photons, even if image infinity time). Durham, NC
[email protected] College of Staten Island Foundation, Inc. Then (optional) you can clean further by using a 70% EtOH wet kimwipe to remove any residual film of oil (you can do this in your lab). There is an channel unmixing "matrix" to let you unmix 'spectral overlaps'. 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). Since we have four laser lines, generally two, three or four scan tracks is sufficient. 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! For more information,visithttp://www2.leica-microsystems.com/e/149501/jhu-lasx-/3tsp5w/258951420, For Confocal applications, call or email Geoff Daniels (senior sales): THis should not be seen as 10x / 2x = 5x advantage, but rather: one time investment ---> 7+ years of user's benefiting from 10x greater throughput (I estimate lifetime of SP8 is 8+ years,could be even longer). Leica DMi8 M System, Do you want help to ensure reliable measurements? Note: HyD's detection must be at least 5 nm from laser lines, for simplicity, for SAFETY please restrict your HyD emission bands to at least 10 nm from any laser line. Microscope Details. Yes, you can acquire an ok image. You increase your sample throughput with the, Choose between manual, coded or automated functions, You can choose between manual or motorized version, Activate with push of 1 button the macro mode to get a quick overview on the sample, Change between contrast mode and magnification simultaneously (e.g. any user who puts a USB stick on an image core acquisition PC will have their USB drive confiscated and tossed int the biosafety trash. Our Lecia SP8 confocal microscope was purchased on a budget, so between the price and capabilities of a 'baby confocal' like Leica SPE, and a 'fully equipped' confocal microscope (example: our SP8 has only 3 detectors, no incubator with 3 detectors, can acquire many channels by sequential scanning multiple tracks). We bill in half hour increments. The HyVolution2 computation (www.svi.nl Huygens deconvolution software with HyD detectors) always generates 16-bit image range (0 ~65535), that is, scaled to the full range (to 65535) even if the photon counts are low (I usually do not use the one PMT on our SP8). Overview. The ISU Advanced Bioimaging Facility is open to users from throughout the Midwest region. The new DMi8 S platform extends the flexibility of the DMi8 microscope, adding high speed control, Infinity TIRF and Infinity Scanner modules, plus advanced software capabilities to create the solution for your advanced live cell imaging. Te Leica SP8 makes it easy to create sequential scan tracks with different pinhole size, so you can test it yourself. Combine high speed External Filter Wheels with multi-position stage experiments and benefit from filter wheels with high precision stage control. we want ACCM staff to be able to use at least 20 of the ~40 hour work week hours). The two Leica HyD detectors are best operated in "counting mode" = photon counting. More Freedom. The Leica SP8 confocal microscope is on a Leica DMi8 inverted microscope stand (DMi8CEL). That is, if imaging air (R.I. 1.0), physical step and "optical step" in the specimen are the same. prior user got oil on the 20x lens), contact George immediately (email above; Ross 913 office [903 during covid-19 space utilization reduction), 305-764-2081 cell) George cleans the lenses. Leica DMi8 C System, Are reproducible image settings and reliable measurements important? The user needs to learn how to use THIS confocal microscope before going on to any highly specialized experiments (i.e. Each user should contact George McNamara (email
[email protected]) the fluorophores and specimens they plan to use. Long term, the SP8 scanhead does not have an X1 port -- this is upgradable in the field. Specifications. Line accumulation number of times each line is scanned I usually recommend starting with 10 (max is 16), which adds up to a good quality image (if specimen has good fluorescence). MicFac -- that have found money for super-resolution. All three models can have X1 port with additional detectors, such as two S dtectors or four APDs (avalanche photodiodes, which have ~80% quantum efficiency in red-NIR but slower photon counting and dynamic range per detector than S detector). The scheduling will move to iLab when the ACCM's iLab page is activated. No messes! The Department of Anesthesiology & Critical Care Medicine (ACCM) users are aiming to use 20 hours per week, that is,50% of JHU business hours access (it's ok if ACCM uses more). Microscope:We acknowledge the Department of Anesthesiology & Critical Care Medicine (ACCM) for access to their Leica SP8 confocal microscope. Web page: http://confocal.jhu.edu/current-equipment/leica-sp8-confocal-microscope, iLab: https://johnshopkins.corefacilities.org/schedules/345585# (JHU login). 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). 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). No USB sticks !!! * Typical immunofluorescence experiment: DAPI, Alexa Fluor 488, Alexa Fluor 568 (or Alexa Fluor 555), Alexa Fluor 647. Refractive index of the Leica immersion oil is 1.518. Find company research, competitor information, contact details & financial data for SARL DEBRUYNE of LAMBERSART, HAUTS DE FRANCE. In practice; good luck getting anywhee close to that resolution on biological specimens and in a building that vibrates (all buildings do). NYU.edu requires JavaScript be enabled in your browser in order to use important features of the site. Ideally purchase FOUR SMD HyD's and negotiate trade-in of the current two 2nd gen HyD's. GM recommends specimen refractive index 1.515 for oil immersion objective lenses (both in calculation and as your mounting medium). Invest in the features you need for your work now and be prepared for future requirements. Swanson Biotechnology Center Reserve theLeica SP8 Confocal MicroscopeRead theUser Manual, 2800 Victory Blvd This new detector is a new design (ask your local Leica rep what is it), with the HyD(TM) now being just a trademark. Probably more important to refractive index match your choice of mounting medium (ex. The Infinity Port Connector, along with complete optomechanical design documentation, opens the Leica DMi8 light path to any accessory you want to add. If you think you have a better way, convince George you are right and we will change. Scanner speed 1 to 1800 Hz, default 400 Hz, typically used at 600 Hz to enable full field imaging (zoom 0.75x). Z all the way down. Nathan Shaner's 6/2019 bioRxiv preprint on AausFP1 bright new GFP could be excited precisely at excitation maximum, AOBS emission side precisely cover emission peak and same for the YFP varient). Note: 0.3 AU and "Area = pi r^2"implies 9% intensity. SP8 offers 0.75x - 40x zoom, so effectively 42x 'total mag' to 'very high' zoom. Environmental Enclosure with heat and humidity. * FALCON = FAst Lifetime CONtrast (Fluorescence Lifetime on HyD photon counting detectors) ==> potential future upgrade of SP8). Get the latest business insights from Dun & Bradstreet. 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). We then pivot to the user specimen(s). Deconvolution -- i.e. If you use #0, ~100 um thick, you can get an additional ~70 um working distance, with only modest loss of image quality from standard coverglass. Define both digital and analog signals, and set up the trigger signaling independently from the image acquisition with exact timings and full reproducibility. Enable "Calculate a Point Spread Function" to have the web page compute and display XY and XZ images of point spread. Analysis of immunofluorescence as well as functional and biochemical kinetic studies can be performed on viable cell samples. This is a state of the science/engineering confocal microscope - we want everyone who can benefit from the LeicaSP8 confocal microscope to be able to get excellent data with it. If dim signal: 16 line accumulation and 'whatever needed' frame accumulation you need to get good data (overnight or over weekend if necessary though you should think about better fluorophores, more laser power, tyramide signal amplification, etc, to get better signal). 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. and finding sources to take care of your data). HyD's in photon Counting mode, usually 10 line accumulation, no frame accumulation. If you are unwilling to follow instructions, go someplace else). No matter what experiments you have in mind, LAS X Navigator is the key to all applications on your DMi8 S platform. Dr. Alessandro Esposito, Hutchison MRC Research Centre, University of Cambridge, UK. Leica SP2 AOBS Confocal Microscope; Leica SP8 Confocal Microscope; . If you have coverglass-slide or 35 mm imaging dish, typically your specimen should look completely transparent (or nearly transparent if thin tissue section or tissue culture cells). so we choose to not provde recommendations for this lens. Information on the advantages on inverted microscopy. POWER HyD(TM) are as fast or faster than 3rd gen HyD, so in fast FLIM mode, a whole lotta data (which 2020 computers can deal with: E-ATX motherboard, PCIE gen4, NVidia Ampere GPUs, 1+ Terabyte fast RAM, lots of PCIe gen 4 NVMe SSD drives (re ASUS Hyper M2 or GloTrends cards), 10Gbe Ethernet (maybe 40Gbe) to local 'distributed computing network'. You can toggle back and forth between "NAV" and TCS SP8 modes (GUI = Graphical User Interface) - most LAS X users will find configuring scan settigns and spectral settigns simpler in the more familiar TCS SP8 "mode". See also their "Explanation: proper images" and additional text on their web page. 0.1 wt% fluorescein sodium salt was added to the PVA precursor as a fluorescent marker, and florescent HA-PVA hydrogels . Switch from searching image by image to seeing the full overview of your samples. You will find a more detailed list of local contacts here. ZERO BIAS - scores, article reviews, protocol conditions and more Combine techniques even further using the fully automated, super-resolution capable Infinity TIRF to analyze membrane dynamics. only fully trained users should touch the instrument and PC. Detailed Specifications Scanhead Andor Dragonfly 505 unit with Borealis illumination spinning disk confocal - 2 pinhole sizes (25 um and 40 um) TIRF microscopy via a separate lightpath Motorized dichroics - BGR (405_13nm/488_13nm/561_6nm/640_16nm) and CY (405_13nm/445_13nm/514_14nm/640_16nm) Additional technology on system: Stay in HyVolution2 mode (unless you really need one of the few features only available in TCS SP8 mode, such as LAS NAVIGATOR stage tiling). For more information or to reserve a demonstration time, Most users should start with simple coverglass-slide preparation. On our SP8, with two HyD detectors and one PMT in the middle, this could be HyD1 500-520nm, PMT 520-525nm (the minimum spectral band), HyD3 525-545nm (I would usually leave PMT off). GM then shows how to acquire Z-series (if user project needs most do). We are happy to answer all your questions and concerns. Fully configurable with manual to motorized components, the DMi8 microscope allows you to build the imaging system for your research and budget needs. ==> Dr. McNamara reviews sessions on the sign-in sheet when doing the 'confirm' step in iLab Organizer. * STELLARIS 8 CW and / or pulsed lasers, optional white light laser (WLL) laser (~440 - ~780nm, see data sheet)and AOBS, TauSense "fast FLIM" (fluorescence lifetime imaging microscopy)AND FALCON"fluorescence lifetime contrast" (fast FLIM, Phasors, other fancy FLIM stuff). Run your experiments up to 5x faster, integrate third party triggered components and synchronize the acquisition with the exposure to minimize photodamage. I described Leica's 3rd generation HyD's above. Prolong Glass) and high NA lens immersion medium on this microscope, Leica oil R.I. 1.518. 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. CMN Core has a new Leica Stellaris 8 Confocal microscope. The DMi8 S is a flexible solution for advanced widefield research. 10); better image quality with more photons (ex. Our 10x lens was not purchased for confocal microscopy. Image dimensions (ex. Combined with an increased focus travel range of 12 mm, choose closed loop focus for the highest repeatability with multi-point time lapse experiments. * Having 3 epi-illumination detectors (2 HyD, one conventional) means maximum 3 fluorescence channels per excitation "scan track". Starting from discovery and analysis of single molecules and culminating in breakthroughs in understanding and treating human health, the key to the next scientific discovery lies in finding the missing links connecting your data. 443-257-2941. Then set up high resolution image acquisition automatically using templates for slides, dishes and multiwell plates. You can also look into DNA-PAINT and using our FISHscope for single molecule localization microscopy. fromhttps://downloads.leica-microsystems.com/Leica%20TCS%20SP8/Brochures/Leica%20TCS%20SP8%20Scan%20Head-Flyer_EN.pdf. Huygens Essential deconvolution (SVI Hyugens Essential intregrated into Leica LAS X software). 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). The modular DMi8 inverted microscope is the heart of the DMi8 S platform solution. Leica DMi8 with manual Transmitted Light method sic microscope settings. . We have "HyVolution2" = Leica SP8 confocal and Huygens Essential. Do you prefer personal consulting? Then add later (which is tedious in LAS X). You can perform Super Resolution, TIRF and several photomanipulation tasks within one time-lapse experiment using up to 5 lasers. 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. Brilliant Violet 421 = BV421, also the "421 component" of the many BV tandems (2 channels). * Two excellent photon counting detectors, HyD1 and HyD3 (second generation hybrid detectors). APDs are 70% quantum efficiency in the red and near infrared (~600 - 900 nm emission), which would enable further multiplexing, discussed at: https://www.linkedin.com/pulse/resolution-blues-meets-21plex-salute-fluorescence-basic-mcnamara, https://www.linkedin.com/pulse/bd-biosciences-listed-tandems-horizon-brilliant-violets-mcnamara. 440 nm 80 MHz, or fiber laser) was purchased, could do "fast FLIM" (would also need upgrade of LAS X software to LAS FALCON fluorescence lifetime contrast). Conventional fluorescence filters for eyepiece vizualization: 4215 French Family Science Center 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. The Leica DMi8 is built with flexibility in mind. 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. Use JHU iLabs ACCM Confocal Microscope for scheduling in advance. GM configures settings, evaluates crosstalk, adjusts any settings if needed, explaining to user, saves settings which user can load for future sessions. June 13, 2018:Leica SP8 is now in Ross S910A(S = Service corridor). I note that MetaMorph can open 16-bit TIFF (and can open an image directly from LAS X's .LIF container file) but MetaMorph does not read HDF5 and does not deal with 32-bit data. Max image resolution 8192 x 8192 pixels (64 megapixels). 16-bit data mode (HyVolution2 Configure settings 16-bit if you leave the default 8-bit, the highest value is 255, which is modest intensity). This tool of choice is suitable for demanding research applications and untrained operators alike. The instrument is equipped with 4 air cooled lasers allowing for excitation wavelengths of 405, 488, 552 and 638 nm. Find company research, competitor information, contact details & financial data for FORESTIA of LAMBERSART, HAUTS DE FRANCE. This simplfies specimen access, whle implying most users will work with fixed specimens on microscope slides or 35 mm imaging dishes. Our SP8 has space for two more HyD's internally. The HyD detectors allow for the use of very low laser power to preserve sensitive samples and allow for repeated scans with minimal fluorescence bleaching. 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. HyD detector(s): photon counting mode please!
[email protected] Sign in first. Change from the macro (35 mm) to nano (200 nm) with only a click. Location: Ernest Mario School of Pharmacy, William Levine Hall, Room 002. If you logged into myJHU and/or MyJHU->Microsoft OneDrive, sign out of each. This novel design facilitates the integration of additional fluorescence light sources and laser systems for advanced applications like. It may be purged by us from Leica PC or our server at ANY TIME. Lasers: 405 nm diode, Argon with 458, 476, 488, 496 and 514 nm lines and a White Light Laser with spectral emission from 470-670 nm. The DMi8 S platform is powered by LAS X Synapse advanced sequencer, which allows you to freely specify the behavior of connections and create fast imaging sequences to analyze an organisms response to external stimuli delivered via third party devices. Danaher Inc andor dragonfly leica dmi8 spinning disk confocal microscope Andor Dragonfly Leica Dmi8 Spinning Disk Confocal Microscope, supplied by Danaher Inc, used in various techniques. 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. If you image through a 400 um thick object object that is R.I. 1.600, the 'apparent Z' is 250 um that is, the Z-motor will travel 250 um. Page 69: Brighteld (Tl) PI-TL" lter cube using LAS X software. Microscope Platform (Leica DMI8 CS, 2023) Fully motorized stage for optimized for tiling and navigation; Super Galvo Z stage (500 nm movement . visithttp://www2.leica-microsystems.com/e/149501/jhu-lasx-/3tsp5w/258951420. "A Zeiss LSM510 Confocal Microscope for Health Sciences Research" 04/01/00-03/31/01 NSF MCB9982161 (PI Wandinger-Ness, A.) We encourage you to go back to your office/lab to export to TIFF and/or JPEG on your own computers (not fill up our PC and server space with your exports). (i) If ACCM users get above 20 hours per work week: great. Staten Island, NY 10314 Below: SVI.nl recommendations for the 63x/1.4NA lens, for Brilliant Violet 421: 405nm excitation, 440 nm emission. The DMi8 S is a flexible solution for advanced widefield research. Bioz Stars score: 86/100, based on 1 PubMed citations. Talk to our experts. * $27/hour for fully trained expert users, during JHMI business hours (Mon-Fri 9am-5pm). Inverted microscopes enable you to save time and money. widefield microscope resolution equation: dxy = 0.61 * Lambda / NA Nyquist theorem (2D) suggests ~3x sampling for pixel size. This enables much simpler quantitation of fluorescence signals than conventional PMTs whose digitizers output values that depend on both the High Voltage ("HV") gain and the "offset". No specimen on microscope, no obstruction over objective lens (ex: make sure slide holder railing not over the lens). The DMi8 modular research microscope is the heart of the DMi8 S system. 500-540nm with adjacent 8nm bands; no need to waste any of the five internal detector positions with a PMT like our SP8 /// I also note that the STELLARIS external X1 port could be configured to be used with POWER detectors, would be great to have say 4 more POWER detectors externally for nine total [or even more with more money). Motorized XY scanning stage (control by remotes or software, do not touch the yellow motors). We have a NAVIGATOR license. Price from $9.99to $1999.99 inverted leica dmi8 sp8 confocal microscope- by Bioz Stars, 2023-05 96/100stars With a standard PMT, it is very difficult to 'translate' the intensity value -- the value depends on the detector gain and offset values (each can be chosen poorly, re https://www.youtube.com/watch?v=VA7J0KkanzM ) -- obtained to either (i) photon counts, (ii) number of fluorophores, or (iii) number of antibody molecules (immunofluorescence) or oligo probes (single molecule RNA FISH. 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. The Leica Application Suite (LAS) software lightens your workload by offering a variety of expert modules, e.g. Tandem scanners (select at software initialization). 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. All lasers to stand by The peak "PQE" ~ 55% (QE accountign for the fill factor of the detector), so higher than 3rd gen HyD (the 'raw' QE would be higher). 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. This confocal laser scanning microscope will provide an advanced multicolor semi-super-resolution imaging system to image fixed or live sample with fast acquisition in 4D. Leica DMi8 inverted microscope with X-Y motorized stage; Metal halide fluorescence lamp & halogen transmission lamp; 10X, 20X Oil, 25X Water, 63X Water DIC & 63X Oil DIC objectives . . **See alsohttps://svi.nl/NyquistCalculatorfor SVI (Huygens) recommendations for optimal XY pixel and Z-step size. The S detectors and HyD are photon countign and 'fast FLIM' capable.