Overview:  The Stellaris 8 is the newest microscope in Leica's SP2, SP5, SP8 series of high-end laser scanning confocal microscopes. It features a redesigned optical path for greater efficiency in light collection and detection compared to previous iterations.

Stellaris features an expanded (compared to SP8) white light laser (WLL) capable of excitation in any wavelength from 440nm to 790nm, and an acousto-optical beam splitter (AOBS) to select/introduce up to eight laser lines (at 8nm intervals) at a time. In addition to the WLL, there is a near-UV 405 laser, so excitation spans the spectrum from UV to deep red. The acousto-optical tunable filters (AOTF) make it possible to detect a wide range of emission wavelengths using any or all of the two chilled HyDXs (single molecule sensitive hybrid GaAsP detectors), two HyDSs (new blue light sensitive, silicon-based detectors unique to the Stellaris) or the HyDR (a new near-IR emission optimized hybrid GaAsP detector unique to the Stellaris). Our system is capable of fluorescence lifetime (FLIM) and tau-sense imaging, giving it the ability to separate dyes based on wavelength and/or fluorescence lifetime. This greatly expands the number of dyes which can be separated in a single sample.Time gated fluorescence detection and tau-sense-mediated separation is possible on all detectors. Spectral and lifetime scanning and unmixing are also available.

Location: KCBD 1250G

Training Contact: Lorraine Horwitz

Fluorophores this microscope can image:

• Violet/near UV (ex: DAPI, Alexa 405)
• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)
• Near IR (ex: Alexa 700, Alexa 750)

Excitation Light Source(s):

• White light laser -- ANY wavelength from 440nm - 790nm. Up to 8 lines at once with 8nm spacing
• Near UV laser at 405nm

Emission Detection:

• Custom emission detection of any wavelength range between 410nm and 850nm
• 2 chilled, single-molecule sensitive hybrid GaAsP/PMT detectors (HyDXs) with optional tau-sense fluorescence lifetime gating
• 1 chilled, single molecule sensitive hybrid GaAsP/PMT detector optimized for near IR detection (HyDR) with optional optional tau-sense fluorescence lifetime gating
• 2 Leica-exclusive, broad dynamic range silicon photon detectors (HyDSs)
• Transmitted light detector with DIC polarizers for 10x, 63x and 100x objectives
• 8-, 12-, or 16-bit grayscale output

Objectives (base magnification, additional optical zoom is possible):

• 10x / NA 0.4 dry 2.74mm WD (506407)
• 16x / 0.6 2.5mm working distance (WD) multi-immersion (water, oil or glycerol) with correction collar rated for BABB / cleared tissue imaging (must be installed before use) (506533)
• 20x / 0.75 dry 0.62mm WD (506517)
• 40x / 1.25 oil 0.24mm WD (506358)
• 63x / 1.4 UV oil 0.14mm WD (506350)
• 100x / 1.45 oil 0.13mm WD (506378)
• 86x / 1.20 water 0.3mm WD (506333)
  (must be installed before use)

Sample Holder:

• Inverted platform for imaging of 1x3 inch slides or 35mm dishes
• Automated XY stage for tiling / multipoint scanning
• Navigator software for non-rectangular tiling. Increases speed for large tiling projects. Includes a fast "stage overview" mode
• Chamber slide users see our chamber slide use warning before starting your cultures!

Special Features:

• DIC optics matching 10x, 63, and 100x objectives
• Extended white light laser for excitation from 440nm-790nm
• Pulse picker on laser to adjust excitation pulse rate and increase likelihood of fluorescence lifetime difference detection
• Extended emission detection range out to 830nm with new HyDR detector optimized for near IR photon capture
• Tau-sense addition to time gates allows for software-assisted separation of dyes based on fluorescence lifetime in addition to wavelength
• FLIM/FALCON software for fluorescence lifetime applications
• Leica AFC (automatic focus correction) uses a laser at 850nm to hold focus over long-term timelapse

Overview: The Leica SP8 is a laser scanning confocal capable of fluorescence lifetime (FLIM), 3-color 3D STED and tau-STED imaging.

It features a white light laser (WLL) capable of excitation in any wavelength from 470nm to 670nm, and an acousto-optical beam splitter (AOBS) to select/introduce up to eight laser lines (at 8nm intervals) at a time. In addition to the WLL, there are Argon and UV lasers for imaging, photobleacing and photoactivation, so exciation spans the spectrum from UV to deep red. There are also three depletion lines for STimulated Emission Depeletion (STED) superresoluion: 775nm (depletes red and far red dyes), 660nm (depletes orange and yellow dyes) and 592nm (depletes green dyes). The acousto-optical tunable filters (AOTF) make it possible to detect a wide range of emission wavelengths with unlimited range on each of three HyDs (hybrid GaAsP detector) two PMTs (photon multiplier tube). Spectral scanning / unmixing in exciation and/or emission is available. Time gated fluorescence detection is possible on the three HyD detectors.

Location: KCBD 1250G

Training Contact: Lorraine Horwitz

Fluorophores this microscope can image:

• Violet/near UV (ex: DAPI, Alexa 405)
• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)
• Near IR (ex: Alexa 700)

Excitation Light Source(s):

• White light laser -- ANY wavelength from 470nm - 670nm. Up to 8 lines at once with 8nm spacing
• Near UV laser at 405nm
• 5 line Argon laser at 458, 476, 488, 496 and 514nm

Lasers for STED depletion:

• pulsed 775nm (depletes red and far red dyes)
• continuous wave 660nm (depletes orange and yellow dyes)
• continuous wave 592nm (depletes green dyes)

Emission Detection:

• Custom emission detection of any wavelength range between 410nm and 800nm
• 2 chilled photon multiplier tubes (PMTs)
• 2 chilled, single-molecule sensitive hybrid GaAsP/PMT detectors (SMD-HyDs) with optional time gating
• 1 non-SMD hybrid GaAsP/PMT detector (HyD) with optional time gating
• Transmitted light detector
• 8-, 12-, or 16-bit grayscale output

Objectives (base magnification, additional optical zoom is possible):

• 10x / NA 0.4 dry wd=2.74mm 506407
• 20x / 0.7 multi-immersion (water, oil or glycerol) wd=0.66mm 506343
• 40x / 1.25 oil wd=0.24mm 506358
• 63x / 1.4 UV oil wd=0.14mm 506350
• 100x / 1.45 oil (STED-rated) wd=0.13mm 506378
• 93x / NA 1.3 glycerol wd=0.3mm 506417 (must be installed prior to use, STED-rated, motorized correction collar)
• 86x / NA 1.20 water wd=0.3mm 506333 (must be installed prior to use, STED-rated, motorized correction collar)

Sample Chamber:

• Inverted platform for imaging of slides or dishes
• Automated XY stage for tiling / multipoint scanning
• "SuperZ" galvo focusing stage, 1.5 mm range. Demonstrated capability of three cell volumes per second (12 x 1-mm optical slices each)
• Navigator software for non-rectangular tiling. Increases speed for large tiling projects. Includes a fast "stage overview" mode
• Chamber slide users see our chamber slide use warning before starting your cultures!

Special Features:

• Capable of Fluorescence Lifetime Imaging (FLIM) and lifetime enhanced STED (tau-STED) imaging
• Capable of 2D or 3D superresolution imaging in XY, XZ or XYZ.
• AOBS (acousto-optical beam splitter) plus sequential scanning capability allows for rapid sequential scanning of fluorophores with minimal bleed-through or cross-talk
• Hybrid GaAsP detectors (HyDs) and chilled SMD HyD detectors feature digital time gating (0.1 nanosecond increments, 3.5 nanosec. (min) to 12 nanosec (max) range) and photon counting mode.
• Transmitted light detector features DIC polarizer/analyzer with prisms for 20x, 40x and 63x.
• Notch filters for 488, 561, 594 and 633 from the WLL
• Spectral scanning across excitation, emission, or both, allowing for separation of fluorophores with similar ranges (e.g. GFP and FITC) through spectral unmixing.
• Time-gating on HyDs allows for differentiation of fluorophores with similar spectra or enhancement of STED resolution from the pulsed 775nm laser.
• Tandem scanner with dual scanning galvanometer mirrors allows for either high speed scanning (max 16,000 Hz scan rate) 25 images/sec at 512x512; strip scans to 333 fps (5 channels) OR high pixel density scanning (imaging to 4k x 4k [16 megapixels] per channel in galvo mode).
• Standard galvo scanner includes beam park for FRAP, bleaching and photoactivaiton
• Optical zoom for sampling to 5nm pixel size
• Wizards for FRAP, FRET and Live Data Mode
• 3D/4D reconstruction software built in to LAS_X image collection software
• LAS_X Leica confocal software on Windows 10
• Off-line version of LAS_X software available on facility workstation

​NOTE: The 592 depletion laser on this system is no longer functional.  We do not have a service contract for this end-of-life system, so we will not be replacing this laser. Anyone in need of confocal imaging with DAPI can use the SP5 2-photon, SP8 or Stellaris.  Anyone in need of super-resolution imaging has likely already moved to the SP8 3 color 3D STED system.  The rest of the system continues to function, so confocal with green, red and far red fluorophores is still possible.

Leica SP5 STED - October 18, 2023 - We swapped this Argon (488nm) laser with the one from the SP5 2-photon which was starting to malfunction. We recommend keeping the power level between standby and 20% (under configurations->laser) to decrease the chances of the laser malfunction and prolong the life of the laser.  We do not know how long this laser will last, but we do know if the malfunction becomes more serious we will have to retire this microscope (see no service contract for this end-of-life system above).

Overview: The SP5 II is an advanced, high speed laser scanning confocal platform. It includes an acousto-optical beam splitter (AOBS) to select/introduce most excitation laser lines. There are eight excitation lines available, spanning the spectrum from green to far red. The acousto-optical tunable filters (AOTF) make it possible to detect a wide range of emission wavelengths with unlimited range on each of three PMTs (photo multiplier tubes), two HyDs (hybrid GaAsP detector) or two APDs (avalanche photodiodes).

Location: KCBD 1250F

Training Contact: Lorraine Horwitz

Fluorophores this microscope can image:

• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)
• Near IR (possible but not ideal, ex: Alexa 700)

Excitation Light Source(s):

• 5 line Argon laser at 458, 476, 488, 496, and 514nm
• DPSS laser at 561nm
• orange HeNe laser at 594nm
• red HeNe laser at 633nm

Emission Detection:

• Custom emission detection of any wavelength range between 410nm and 800nm
• 3 chilled photon multiplier tubes (PMTs)
• 2 hybrid GaAsP/PMT detectors (HyDs) with optional time gating
• 2 internal avalanche photodiode detectors (APDs) for high sensitivity (green/red and CFP/YFP filters available)
• Transmitted light detector with DIC polarizer/analyzer plus prisms for most objectives available
• 8-, 12-, or 16-bit grayscale output

Objectives (base magnification, additional optical zoom is possible):

• 10x / NA 0.4 dry wd=2.74mm
• 20x / 0.7 multi-immersion (water, oil or glycerol) wd=0.26-0.17mm
• 40x / 1.25-0.75 oil wd=0.22mm
• 63x / 1.4-0.6 UV oil wd=0.14mm
• 100x / 1.40 oil (STED-rated) wd=0.13mm
• 50x / 0.9 dry wd=0.28mm (must be installed prior to use)
• 63x / 1.3 glycerol CORR wd=0.30mm (must be installed prior to use)

Sample Chamber:

• Full wrap incubator box with warm air heating for live samples
• Inverted platform for imaging of slides or dishes
• Automated XY stage for tiling / multipoint scanning
• "SuperZ" galvo focusing stage, 1.5 mm range. Demonstrated capability of three cell volumes per second (12 x 1-mm optical slices each)
• Chamber slide users see our chamber slide use warning before starting your cultures!

Special Features:

• Continuous wave depletion laser at 592nm allows for single or dual color super resolution (STED method) with either the resonant (high speed) or galvo (high pixel density) scanners.
• STED mode allows for resolution of particles down to 50nm FWHM (cyan, green, yellow fluorophores only)
• Tandem scanner with dual scanning galvanometer mirrors allows for either high speed scanning (max 16,000 Hz scan rate) 25 images/sec at 512x512; strip scans to 333 fps (5 channels) OR high pixel density scanning (imaging to 8k x 8k [64 megapixels] per channel).
• Standard scanner includes beam park for FRAP, bleaching and photoactivaiton
• Sequential scanning capability allows for rapid sequential scanning of fluorophores with minimal bleed-through or cross-talk
• Wizards for FRAP and FRET
• AOTF (acousto-optical tunable filters) for spectral scanning, allowing separation of fluorophores with similar ranges (e.g. GFP and FITC) through spectral unmixing
• LAS_AF Leica confocal software on Windows
• Off-line version of LAS_AF software available on facility workstation

NOTES: The chiller for the MaiTai multiphoton laser has FAILED therefore the MaiTai is currently out of service. The rest of the Leica SP5 2-photon microscope is working normally for now, so if your project does not require excitation at longer wavelengths (700nm+) it can continue. This includes intravital imaging without the multiphoton laser.

Leica swapped the Argon laser with the one on the SP5 STED. We recommend keeping the power level low (between standby and 20% under configurations->laser) to prolong the life of the laser.

The HyD2 detector is failing, signal detected is very low. We recommend avoiding this detector if possible.

Overview: This laser scanning confocal system features software selectable conventional or high-speed resonance scanner galvanometer system with three (chilled, high sensitivity) internal PMT detectors (spectral) and two external (NDD) PMT detectors. Six visible laser lines and a tunable NIR pulsed laser (Spectra Physics Mai Tai broadband 710-990nm) provide excitation. The system features turnkey operation and full software control in addition to the features listed below.

Location: KCBD 1250B

Training Contact: Lorraine Horwitz

Fluorophores this microscope can image:

• Blue (ex: DAPI, Alexa 405)
• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)
• Near IR (possible but not ideal, ex: Alexa 700)
• 2-photon imaging of Blue, Cyan, Green, Yellow, Orange - NO LONGER IN SERVICE

Excitation Light Source(s):

• near UV laser at 405nm
• 5 line Argon laser at 458, 476, 488, 496 and 514nm
• HeNe laser at 561nm
• HeNe laser at 594nm
• HeNe laser at 633nm
• MaiTai laser tunable from 710 - 990nm - NO LONGER IN SERVICE

Emission Detection:

• Custom emission detection of any wavelength range between 410nm and 800nm
• 3 chilled photon multiplier tubes (PMTs)
• 2 hybrid GaAsP/PMT detectors (HyDs)
• 2 non-descanned detectors (NDDs)
• Transmitted light detector
• 8-, 12-, or 16-bit grayscale output

Objectives (with additional optical zoom possible):

• 20x / NA 0.8 dry
• 40x / 0.8 water
• 63x / 0.9 water
• 63x / 1.4 oil
• 100x / 1.46 oil
• 25x / NA 0.95 water (must be installed)
• Objectives listed under the SP5 STED description (must be installed)

Sample Chamber:

• Custom fit full incubation jacket (clear) for increased thermal stability during live cell experiments
• Inverted platform for imaging of slides or dishes
• LSM objective inverter device (allows system to be used as if it were an upright microscope platform)
• Automated XY stage for tiling / multipoint scanning
• "SuperZ" galvo focusing stage, 1.5 mm range. Demonstrated capability of three cell volumes per second (12 x 1-mm optical slices each)
• Chamber slide users see our chamber slide use warning before starting your cultures!

Special Features:

• FRET, deconvolution, and 3D software wizards
• Automated notch filters
• Spectra Physics Mai Tai broadband Ti:sapphire laser with 10W pump (tunable from 710 to 990 nm) (up to 2W output at 810 nm) - NO LONGER IN SERVICE
• Dual scanning galvanometer mirrors (max 16,000 Hz scan rate) 25 images/sec at 512x512; strip scans to 333 fps (4 channels). Standard scanner with beam park, bleaching, imaging to 8k x 8k (64 megapixels) per channel.
• AOTF control of visible laser lines, EOM and ND filter attenuation of NIR
• Excitation and emission scanning capability with linear unmixing
• Automatically optimized confocal pinhole apertures
• Transmitted detector with LWD and oil lenses and filters suited for SHG imaging

Overview: The Caliber I.D. RS-G4 is an upright laser scanning confocal microscope with a motorized XY-Z stage and software designed for large format (tiled) images. This system is ideal for high speed, high quality tiling in 2D or 3D and will handle samples up to 120mm x 80mm x 6mm thick.

Location: KCBD 1250F

Training Contact: Christine Labno

Fluorophores this microscope can image:

• Blue (DAPI, Alexa 405)
• Cyan (CFP)
• Green (GFP, Alexa 488)
• Yellow (mCitrine, YFP)
• Orange (Cy3, Alexa 543)
• Near Red (Texas Red, Alexa 594)
• Far Red (Cy5, Alexa 647)
• Near IR (Alexa 700)

Excitation light source(s):

• Near UV laser at 405nm
• solid state laser at 488nm
• solid state laser at 561nm
• solid state laser at 640nm
• solid state laser at 785nm

Emission detection:

• 2 photon multiplier tubes (PMTs) for emission detection
• Seven band pass filters for emission range selection: 450/70, 520/44, 550/88, 600/52, 630/69, 670/30 and 832/37
• 16-bit tif or Imaris .ims file output.

Objectives (base magnification, additional optical zoom is possible):

• Supports dry, oil and water immersion objectives with a wide range of magnifications available (2x - 100x).
• Objectives must be programmed in prior to use.

Sample Chamber:

• Upright platform for imaging of slides
• Imaging in dishes limited, only possible with dipping (coverslip-free) objectives
• Fast ribbon / strip mosaic scan mode plus conventional tiling mode, both with real time / online stitching

Special Features:

• 8kHz Resonant-scanning confocal
• Large mosaic creation up to 120 x 80 mm with high precision scanning stage.
• Fluorescence and/or reflectance confocal imaging
• Automated scanning of single, simultaneous pairs or sequential channels.
• Automated collection of time-lapse, X-Y-Z, multi-stage points, multiple wavelengths and snapshots.
• 1024 x 1024 pixel resolution

Overview: Fully automated, inverted, Yokogawa-type spinning disk confocal ideal for imaging live samples. Features an automated XY stage and piezo-controled fine Z stage (focus) positioning for multi-point scanning plus incubator box for temperature, CO2 and humidity control. Slidebook software controls filter cube turret, objective turret, and high-speed brightfield shutter. SoRa with microvolution for nearly instantaneous deconvolution is ideal for subdiffraction (2-fold improvement over confocal) live cell imaging with little phototoxicity.

Location: KCBD 1250F

Training Contact: Khalil Rodriguez

Fluorophores this microscope can image:

• Violet/near UV (ex: DAPI, Alexa 405)
• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)

Excitation Light Source(s): 

• Near UV laser at 405nm
• Solid state laser at 445nm
• Solid state laser at 488nm
• Solid state laser at 561nm
• Solid state laser at 638nm

Emission Detection:

• Two Prime 95B Back Illuminated Scientific CMOS (11x11um square pixels @ 18.7mm field of view)
• blue (DAPI) band pass emission filter at 445/58
• band pass filter for cyan
• green (FITC, GFP) band pass emission filter at 525/50
• red (TRITC, mCherry) band pass emission filter at 600/50
• far red (Cy5) band pass emission filter at 706/95

Objectives:

• 10x / NA 0.3 dry (Plan-Neofluar) working distance: 5.2mm
• 40x / NA 1.3 oil (Plan-Apochromat) working distance: 0.22mm
• 63x / NA 1.4 oil (Plan-Apochromat) working distance: 0.14mm
• 100x / NA 1.46 oil (Alpha Plan-Fluar) working distance: 0.11mm

Sample Chamber:

• Inverted platform for imaging of slides or dishes
• OKO full environmental control chamber (constant temperature, humidity and CO2)
• Motorized XY stage for multipoint timelapse and tiling
• Piezo controlled fine Z-stage positioning for 3D imaging

Special Features:

•  Super-Resolution through Optical Reassignmnet (SoRa) is ideal for super-resolution live cell imaging and low phototoxicity.  Specifications of SORa imaging plus deconvolution give approximately 2 fold increase in resolution (from 200nm to 100nm).
• CSU-W1 Yokogawa spinning disk allows for high speed imaging up to 200fps, wide field of view 16mm x 17mm, 25µm and 50µm pinhole disks for lower and higher magnification objectives
• Microvolution GPU accelerated Deconvolution Module for nearly instantaneous deconvolution.
• Options for split-view imaging, NIR imaging, illumination field flattening and super-resolution imaging
• Fast shutter speeds and channel switching for high speed imaging
• Vector high-speed point scanner for photoactivation/photoablation and FRAP experiments
• Full microscope automation through Slidebook software

Overview: Fully automated, inverted, Yokogawa-type spinning disk confocal ideal for imaging live samples. Features an automated XY stage and piezo-controlled fine Z stage (focus) positioning for multi-point scanning plus incubator box for temperature, CO2 and humidity control. Slidebook software controls filter cube turret, objective turret, condenser, DIC and brightfield optics and high-speed brightfield shutter.

Location: KCBD 1250F

Training Contact: Khalil Rodriguez

Fluorophores this microscope can image:

• Violet/near UV (ex: DAPI, Alexa 405)
• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)

Excitation Light Source(s):

• Near UV laser at 405nm
• Solid state laser at 445nm
• Solid state laser at 488nm
• Solid state laser at 561nm
• Solid state laser at 638nm

Emission Detection:

• Two Photometrics Evolve- backthinned, air chilled (-85C) EM-CCD cameras (512x512 16um pixels)
• DualCam DC2 beam splitter for simultaneous imaging of two channels (fliters for CFP/YFP and green/red)
• band pass filter for blue
• band pass filter for cyan
• band pass filter for green
• band pass filter for red
• band pass filter for far red

Objectives:

• 20x / NA 0.5 dry (Plan-Neofluar)
• 40x / NA 1.3 oil (Plan-Apochromat)
• 63x / NA 1.0 water dipping (Plan-Apochromat)
• 100x / NA 1.45 oil (Alpha Plan-Fluar)

Sample Chamber:

• Inverted platform for imaging of slides or dishes
• OKO full environmental control chamber (constant temperature, humidity and CO2)
• Motorized XY stage for multipoint timelapse and tiling
• Piezo controlled fine Z-stage positioning for 3D imaging

Special Features:

• Sperical Aberration correction (SAC) optics for sharp images through thicker specimens
• Fast shutter speeds and channel switching for high speed imaging
• Vector high-speed point scanner for photoactivation/photoablation and FRAP experiments
• Full microscope automation through Slidebook software

​Overview: The Olympus "live cell" DSU can be used for either widefield or confocal fluorescence capture, and since the entire field of view is illuminated at once, z-stack and time lapse capture can be faster than on the laser scanning confocals. The live cell DSU is on an inverted platform and features water immersion objectives, a motorized stage and filters for a broad range of fluorophores, from blue to far red, plus DIC optics.

Location: KCBD 1250F

Training Contact: Christine Labno

Fluorophores this microscope can image:

• Violet/near UV (ex: DAPI, Alexa 405)
• Cyan (ex: CFP)
• Green (ex: GFP, Alexa 488)
• Yellow (ex: mCitrine, YFP)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)

Excitation Light Source and Filters:

• Lumencore SOLA Fish LED light engine for even, flicker-free illumination
• blue/violet excitation filter at 350/50
• blue excitation filter at 405/12
• cyan excitation filter at 436/10
• green excitation filters at 480/25 and 490/20
• yellow excitation filter at 500/20
• orange excitation filter at 555/28
• red excitation filter at 565/25
• far red excitation filter at 635/20

Emission Detection:

• Photometrics Evolve- EMCCD camera (512 x 512, 16um pixels)
• blue emission filter at 457/50nm
• cyan emission filter at 470/30
• green emission filters at 525/40 and 528/38
• yellow emission filter at 535/30
• red emission filters at 617/73 and 620/60
• far red emission filter at 685/40

Objectives:

• 10x / NA 0.3 dry (UPlanFlN, 10mm WD)
• 10x / 0.4 water (UPlanApo)
• 20x / 0.7 water (UApo340, 0.35mm WD)
• 40x / 1.15 water (UApo340, 0.25mm WD)
• 60x / 1.2 water (UPlanSApo, 0.28mm WD)
• 100x / 1.45 oil TIRFM (PlanApo)
• 2x / 0.06 dry (must be installed prior to use)
• 4x / 0.16 dry (must be installed prior to use)
• 40x / 0.6 dry LWD (must be installed prior to use)
• 150x / 1.45 oil (must be installed prior to use)

Sample Chamber:

• Fully automated, inverted platform (model IX81) for imaging on slides, chamber slides, 35mm dishes and multi-well plates (see our guidelines on use of chamber slides and well plates)
• Heated stage platform and micro chamber available for live cell preps (35mm dish configuration)
• Motorized xy-galvo stage for imaging multiple areas per dish and automated tiling of large specimens
• Chamber slide users see our chamber slide use warning before starting your cultures!

Special Features:

• Image capture through SlideBook software
• 3D and 4D (volume over time) rendering through SlideBook
• 100% - 1.5% neutral density filters available
• Interchangable pinhole disks for many magnifications (special request only)
• Dual cube viewing through the occulars (blue/green, green/red and red/far red)

Overview: NO LONGER IN SERVICE, information here is for legacy/publication generation purposes. The Olympus "fixed cell" DSU is one of two Olympus spinning disk confocal systems available in the facility. These systems can be used for either widefield or confocal fluorescence capture, and since the entire field of view is illuminated at once, z-stack and time lapse capture can be faster than on the laser scanning confocals. The fixed cell DSU features high NA oil objectives, high sensitivity camera and filters for a broad range of fluorophores, from blue to far red, plus DIC optics.

Location: NO LONGER IN SERVICE, information here is for legacy/publication generation purposes

Training Contact: NO LONGER IN SERVICE, information here is for legacy/publication generation purposes

Fluorophores this microscope can image:

• Blue (ex: DAPI, Alexa 405)
• Green (ex: GFP, Alexa 488)
• Orange (ex: Cy3, Alexa 543)
• Near Red (ex: Texas Red, Alexa 594)
• Far Red (ex: Cy5, Alexa 647)
• Near IR (ex: Alexa 700)

Excitation Light Source and Filters:

• Lumencore SOLA Fish LED light engine for even, flicker-free illumination
• blue/violet excitation filter at 387/11
• green excitation filter at 485/20
• reds excitation filter at 560/25
• far reds excitation filter at 650/13
• near IR excitation filter at 710/40

Emission Detection and Filters:

• Photometrics Prime 95B 95% QE sCMOS camera (1200 x 1200 11um pixel size)
• blue emission filter at 440/40
• green emission filter at 525/30
• red emission filter at 607/36
• far red emission filter at 684/24
• near IR red emission filter at 775/46

Objectives:

• 10x / NA 0.3 dry (UPlanFlN,10mm WD)
• 20x / 0.5 dry (UPlanFl, 2.1mm WD)
• 40x / 1.3 oil (Zeiss PlanNeoFl, 0.21mm WD)
• 60x / 1.4 oil (PlanApo)
• 60x / 1.2 water
• 100x / 1.35 oil (UPlanApo)
• 2x / 0.06 dry (must be installed prior to use)
• 4x / 0.16 dry (must be installed prior to use)
• 40x / 0.6 dry LWD (must be installed prior to use)
• 150x / 1.45 oil (must be installed prior to use)

Sample Chamber:

• Inverted platform (model IX81) for imaging through slides, dishes or plates
• z-galvo stage for Z-sectioning

Special Features:

• Image capture through SlideBook software
• 3D and 4D (volume over time) rendering through SlideBook
• Neutral density filters available for long-term time lapse imaging
• DIC polarizer/analyzer plus appropriate prisms for most objectives