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Confocal Microscopes

Cancer & Cardiovascular Research Building (CCRB)

Caliber I.D. RS-G4 Ribbon Scanning Confocal

RSG4

Location: 1-220A CCRB

The RS-G4 has been designed with a compact, high-speed scan head with macro- and micro- imaging optics that allow acquisition of in-vivo and ex-vivo images from slides to fixed/cleared whole tissue mounts in micrometer to centimeter scales using the same instrument.

Learn More About the Caliber I.D. RS-G4 Ribbon Scanning Confocal

In addition, ex-vivo tissue sections can be imaged in both reflectance and fluorescence modes. 360-degree rotation of the scan head allows greater flexibility and precise angling for acquiring images of large tissue, organs or whole animals. This instrument is essentially agnostic as to what objectives can be used. We routinely use our Nikon 10x, 0.5na, 5.5mmWD and the 20x, 1.0na, 8.2mmWD glycerol immersion objectives for imaging of fixed, cleared tissues.

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Equipment Specs

Almost any objective can be used on this scope.  We have options from 1x to 100x, including the new Nikon 20x, 1.0na, 8.2mm WD objective for fixed cleared samples. Resolution is determined by which objective is used with individual frames (1024 x 1024 pixels) captured at up to 25fps.

Highlights

  • Supports standard dry, oil and water immersion objectives
  • Fast, large-scale mosaic creation up to 120 x 80 mm
  • To-the-pixel image accuracy for tiling
  • Automated simultaneous and sequential reflectance and fluorescence imaging
  • Intuitive microscope control taught by trained UIC staff
  • Experiment acquisition setup for automated collection of snapshots, time, X-Y-Z, multi-stage points, wavelength
 
Objectives: 
Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
2x 0.1 PlanApo   ∞/--   Air
4x 0.2 PlanApo   ∞/-- 15.7 Air
10x 0.5 PlanApo Correction Coller   5.5 Glycerol+
10x 0.45 PlanApo DIC N1 ∞/0.17 4 Air
10x 0.3 PlanFluor DIC L ∞/0.17 16 Air
20x 1 PlanApo Correction Coller ∞/0 8.2 Glycerin+
Lasers: 
  • Operating Wavelengths include: 405, 488, 561, 638, 785 nm

Nikon A1R FLIM Confocal Microscope

A1RFLIM

Location: 1-220H CCRB

Features include Nikon's patented PFS Perfect Focus III focus lock technology. LED Illumination systems for both transmitted and reflected light applications. New Lambda coat anti-reflective coated objectives at 4x, 10x, 20x, 20x water immersion, 40x oil, and 60x oil magnifications for DIC. Fluorescence illumination and filters for widefield applications for DAPI, CFP, GFP, YFP, RFP, and Cy5. The scope is equipped for widefield imaging using a Hamamatsu Flash 4 V3 CMOS camera.

  • A1R GaAsP Confocal System
  • Laser Selection: 405nm, 445nm, 488nm, 514nm, 561nm and 640nm.
  • AR1 Confocal Scanner: Nikon's exclusive hybrid scanner allows flexible switching or simultaneous use of two galvo scanners (resonant and non-resonant) this allows ultrafast imaging (up to 420fps) and photo activation imaging required to unveil cell dynamics and interaction.
  • Detectors: Included are 4 confocal detectors (2 GaSaP) plus transmitted detector.

The Picoquant Fluorescence Lifetime Imaging Microscopy (FLIM) system has 405, 485 and 560 nm pulsed lasers and can perform many FLIM and FCS techniques.

Learn more about the Nikon A1R FLIM Confocal Microscope

  • Six (6) laser lines of excitation
  • A system fully configured for screening of plates and slides
  • A modular system for assay development (Nikon Elements and JOBS)
  • Confocal imaging for 3D samples
  • Environmental control for live cell imaging

References:

 Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
4x 0.2 PlanApo   ∞-- 20 Air
10x 0.45 PlanApo   ∞/0.17 4 Air
20X 0.75 PlanApo DIC N2 ∞/0.17 1 Air
20x 0.95 PlanApo Lambda with correction collar LWD ∞/0-2 0.8 Water Immersion
40x 1.3 PlanFluor DIC N2 ∞/0.17 0.24 Oil
40x 0.6 SPlanFluor ELWD, NAMC3 ∞/0-2 3.6-2.8 Air
60x 1.4 Plan DI ∞/0.17 0.13 Oil
60x 1.2
Plan Apo VC with correction collar
  ∞/0.15-0.18 0.27 Water Immersion

* these objectives have a correction collar for various coverslip thickness. See staff for approprate adjustments.

Lasers:

  • 405nm
  • 442nm
  • 488nm
  • 514nm
  • 561nm
  • 640nm

Pulsed FLIM lasers

  • 405nm
  • 470nm
  • 561nm
Detector: 

Two (2) GaSaP  Detectors

Two (2) enhanced PMT detectors

One (1) transmitted light detector

One (1) Hammamatsu Flash 4.0 V3 CMOS camera

 

 

Nikon C2 Upright Spectral Confocal and Widefield Imaging System

NikonC2

Location: 1-220B CCRB

Features include Nikon’s newest motorized Ni microscope body. The newly developed CFI Plan Apochromat λ (lambda) series objectives are the key to the series’ optical performance. Nano Crystal Coat, with its ultra low refractive index, provides brighter, high-resolution and high-contrast microscopy images. The CFI 60 objectives for the Ni system are 10x, 20x, 40x oil, and 60x oil, all with DIC capabilities. An LED illumination system for fluorescence, and filters for widefield using DAPI, GFP, and Tex Red filters. The Ni stand uses Nikon’s proprietary stratum structure, used and highly acclaimed for inverted research microscopes, is now incorporated in an upright microscope, allowing a combination of components to be installed. There is a Prior motorized x/y stage and color camera for tiling of large areas of interest in brighfield, DIC, fluorescence and confocal modes.

  • There are 405nm, 488nm, 561nm and 640nm laser lines for excitation.
  • Dual GaSP detectors with continuous filters for spectral imaging.
  • Three (3) photo-multiplier tubes (PMTs), plus transmitted light detectors.
  • Color digital camera for widefield image capture, including tiling in brightfield, DIC, DAPI, AF488, AF568 and CY5 fluorescence. 
  • Includes Elements modules for 2D and 3D Tracking and Measurements, Ratio and FRET imaging.

Learn more about the Nikon C2 Upright Spectral Confocal and Widefield Imaging System

Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
4x 0.13 PlanFluor PhL DL ∞/1.2 16.5 Air
10x 0.45 PlanApo DIC N1 ∞/0.17 4 Air
20x 0.75 PlanApo DIC N2 ∞/0.17 1 Air
40x 1.3 PlanFluor DIC N2 ∞/0.17 0.24 Oil Immersion
60x 1.4 PlanApo DIC N2 ∞/0.17 0.14 Oil Immersion
100x 1.4 PlanApo VC DIC N2 ∞/0.17 0.13 Oil Immersion

Transmitted Light Filters:

  • ND2, ND4, ND8, and ND16
  • Green and cool blue

Lasers:

405
488
561
647

Detector:

Three (3) PMTs and a transmitted light detector
Nikon DS-U3 color camera

Leica SP5 Multiphoton Upright Confocal

Leica SP5

Location: 1-250D CCRB

Leica DM6000 CFS upright multiphoton microscope equipped with a TCS SP5 MP laser confocal with both galvonomic and resonance scanners and with internal spectral HyD detectors. 405, 458, 476, 488, 514, 543, 594, and 640 nm single photon laser lines and a Mai Tai eHP DS 690-1040 nm tunable multiphoton laser along with 4-channel non-descanned detectors. Objectives available include 63x/NA 1.3 oil and 25x/NA 0.95 water (optimal for intravital deep tissue imaging (up to 2.5 mm)).

Learn more about the Leica SP5 Multiphoton Upright Confocal

Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
5x 0.15 HCX PL FLUOTAR/CS   ∞/-/C 13.7 Air
10x 0.4 HC PL APO/CS Confocal Series ∞/0.17/A 2.2 Air
20x 0.7 IMM CORR/CS Confocal Series ∞/-/C 0.26 Water-Glycerol-Oil
L 25x 0.95 HCX IR APO/CS VISIR ∞/0.17/0 2.4 Water Immersion
L 25x 0.95 HC FLUOTAR VISIR ∞/0/0 2.5 Water Dipping
40x 1.3 HCX PL APO/CS Confocal Series ∞/0.17/0 0.24 Oil
63x 1.4-0.60 HCX PL APO/CS Confocal Series ∞/0.17/E 0.14 Oil

Jackson Hall

Nikon A1RHD Multiphoton Upright Confocal

Nikon A1RMP HD

Location: 1-166B Jackson Hall

Nikon A1RMP confocal is connected to an upright Nikon FN1 microscope featuring a Plan Apo LWD 25x water-immersion, NA 1.1 objective most suitable for imaging deep (it has a 2mm working distance) into a fixed or living sample. In addition to conventional and resonant visible laser confocal imaging, a Spectra Physics 15W Mai Tai eHP tunable IR laser and 4 non-descanned GaAsP detectors allow multiphoton visualization of multiple fluorophores located deep within the specimen. Visible lasers include 405, 457, 488, 514, 561 and 637 nm lines. The system has both galvonomic (4K) and resonance (720fps) scanners that can be used in concert simultaneously for photoactivation and MP imaging with enhanced hybrid descanned PMTs. An ASI piezo motorized stage(link is external) can be used for tiling or multiple-position imaging.

Learn More About Nikon A1RHD MP Confocal

The system is also equipped with Second Harmonic Generation (SHG) detection.  SHG doesn't involve the excitation of molecules like other techniques such as fluorescence microscopy. See UIC staff for training and questions.

This instrument was funded in 2012 by the Minnesota Partnership in conjunction with the Mayo Clinic Foundation, Rochester, MN.

In October 2018 the system was upgraded to the an A1RHD with 1024 resonant scanning and improved signal to noise imaging in resonant mode.

Objectives:

Magnification  NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
10x 0.3 PlanFluor DIC L ∞/0.17 16 Air
10x 0.3 Fluor DIC L ∞/0 2 Water Dipping
10x 0.5 Plan Apo Correction Collar ∞/- 5.5 Glycerol
20x 0.45 PlanFluor DIC L ∞/0-2 7.4 Air
20x 0.75 PlanApo lambda DIC N2 ∞/0.17 1 Air
25x 1.1 ApoLWD DIC 2, Correction Collar ∞/0-0.17 2 Water Dipping
40x 0.8 Fluor DIC M ∞/0 2 Water Dipping
100x 1.3 PlanFluor   ∞/0.17 0.2 Oil Immersion

Nikon A1Rsi Confocal with SIM Super Resolution

A1RsiSIM

Location: 1-159 Jackson Hall

Nikon Ti-E Motorized Microscope featuring Nikon’s patented PFS Perfect Focus III focus lock technology. LED Illumination systems for both transmitted and reflected light applications. New Lambda coat anti-reflective coated objectives at 10x, 20x, 40x, 60x, and 100x magnifications for DIC. Fluorescence accessories for widefield applications for DAPI, GFP, RFP, and Cy5. The Nikon Elements software has the JOBS package that can automate image collection for high content screening of plates.

SIM Structured Illumination System-Using high frequency Structured Illumination, the Nikon N-SIM can achieve image resolution of 85nm, which was previously considered impossible with optical microscopes. Furthermore, with temporal resolution of up to 0.6 sec/frame, N-SIM enables super resolution time-lapse imaging capture of dynamic molecular interactions in living cells.

Super Resolution References

Learn more about the Nikon A1Rsi Confocal with SIM Super Resolution

Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
10x 0.45 PlanApo DIC N1 ∞/0.17 4 Air
20x 0.75 PlanApo VC DIC N2 ∞/0.17 1 Air
40x 0.6 S PlanFluor DIC N1, Correction Collar ∞/0-2 3.6 Air
60x 1.27 PlanApo IR with correction collar DIC N2 ∞/0.15-0.19 0.18-0.16 Water Immersion
60x 1.2
Plan Apo VC with correction collar
  ∞/0.17 0.31 Water Immersion
60x 1.4 Plan Apo Lambda DIC N2 ∞/0.17 0.13 Oil Immersion
40x 0.95 PlanApo DIC MN 2, Correction collar ∞/0.11-0.23 0.14 Air
100x 1.49 SR ApoTIRF with correction collar DIC N2 ∞/0.13 0.12 Oil Immersion

Transmitted Light Filters:

  • ND2, ND4, ND8, and ND16
  • Green and cool blue

Lasers (A1R)

  • 405
  • 488
  • 561
  • 637

Lasers (N-SIM)

  • 405
  • 488
  • 561
  • 647

Detector:

A1R has 3 of 4 GaAsP detectors that supply 20 percent more quantum efficiency. There is also a transmitted light detector and 32-channel spectral detector.

The N-SIM
Andor iXon Ultra 897(link is external) camera

Olympus FluoView FV1000 BX2 Upright Confocal

BX2

Location: 1-162 Jackson Hall

Olympus FluoView FV1000 is designed for high-resolution, confocal observation of both fixed and living cells. The FV1000 offers advances in confocal system performance while providing the speed and sensitivity required for live cell imaging, with minimal risk of damage to living specimens. The system has four fluorescence detectors plus a transmitted light detector for DIC. 

Learn more about the Olympus FluoView FV1000 BX2 Upright Confocal

  • FluoView 1000, Olympus Corporation, USA
  • Automated upright  BX61 microscope base
  • 100W Mercury fluorescent lamp
  • 100W halogen transmitted illuminator with detector
  • PRIOR ProScanII motorized XY stage 
  • DIC optics

Methods Include:

  • Fluorescence
  • Brightfield
  • DIC
  • FRAP
  • FRET

Automated Capabilities:

  • Time-lapse
  • Multi-channel fluorescence
  • Co-localization
  • Z-stacks 

Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
4x 0.16 UPlanApo -- ∞/0.17 13 Air
10x 0.4 UPlanApo BX2-DIC10 ∞/0.17 3.1 Air
20x 0.85 UPlanSApo BX2-DIC20 ∞/-/FN26.5 0.2 Oil Immersion
40x 1.3 UPlanFLN BX2-DIC40 ∞/0.17/FN26.5 0.2 Oil Immersion
60x 1.42 UPlanApo N BX2-DIC60 ∞/0.17/FN26.5 0.15 Oil Immersion
60x 0.9 LUMPlanFL   ∞/0 2 Water Dipping

Filters:

Filter Wavelengths
Excitation Filter (nm) Dichoric Mirror (nm) Emission Filter (nm)
DAPI D350/50 BS20/80 460/50
FITC D480/30 DM405/488 D535/40
TRITC D540/25 DM405/488/543/635 D605/55
Cy5 D620/20 DM458/515 D700/75
Channels Dichoric Mirror (nm) Barrier Filters (nm)
Channel 1 SDM560, SDM510, SDM490 BA505-525, BA480-495, BA430-470
Channel 2 SDM560 BA505-605, BA560-620, BA535-565, BA505-525
Channel 3 SDM640 BA560-660
Channel 4 -- BA655-755
Lasers: 
  • 405nm 25mW Diode
  • 458, 488, 515nm 30mW Argon
  • 543nm 1mW HeNe green
  • 635nm 10mW HeNe red
Detector: 
  • 4 photomultiplier tubes channels
  • 1 transmission detector

Olympus FluoView FV1000 IX2 Inverted Confocal with FLIM detector

IX2

Location: 1-158 Jackson Hall

This system is equipped with pulsed 405 and 485 nm lasers for FLIM. FLIM is the acronym for "Fluorescence Lifetime Imaging Microscopy" and is the technique that maps the spatial distribution of fluorescence lifetimes within microscopic images of fixed or living cells. The fluorescence lifetime measures the time that a population of fluorescent molecules spends in the excited state. The fluorescence lifetime does not change upon intensity variations and therefore lifetime measurements are not dependent on the local concentration of fluorophores, or variations in the optical path of the microscope, the local excitation light intensity, or on the local fluorescence detection efficiency.

The FLIM is a time domain system, and uses pulsed lasers at 405 and 488 nm. The detector on the system is a Hybrid GaAsP cathode with three possible emission filter setups - a 480/30nm band pass for CFP emission, a 460LP and a 525/50 nm bandpass filter for GFP. 

learn more about the Olympus FluoView FV1000 IX2 Inverted Confocal with FLIM detector

Automated Capabilities:

  • Time-lapse
  • Stitching
  • Multi-Channel fluorescence
  • Co-localization
  • Spectral unmixing
  • Z-stacks

External FLIM

  • PicoQuant pulsed lasers at 405 and 485 nm
  • Software includes Becker&Hickl SPCM and SPC Image

Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
4x 0.16 UPlanSApo -- ∞/-/FN26.5 13 Air
10x 0.4 UPlanSApo IX2-DIC10 ∞/0.17/FN26.5 3.1 Air
20x 0.85 UPlanSApo IX2-DIC20 ∞/-/FN26.5 0.2 Oil Immersion
40x 1.3 UPlanFLN IX2-DIC40 ∞/0.17/FN26.5 0.2 Oil Immersion
60x 1.42 UPlanApo N IX2-DIC60 ∞/0.17/FN26.5 0.15 Oil Immersion

Filters:

Filter Wavelengths
Excitation Filter (nm) Dichoric Mirror (nm) Emission Filter (nm)
DAPI D350/50 BS20/80 460/50
FITC D480/30 DM405/488 D535/40
TRITC D540/25 405/488/543 D605/55
Cy5 D640/20 DM458/515 D680/30
Channels Dichoric Mirror (nm) Barrier Filters (nm)
Channel 1 SDM560, SDM510, SDM490 2nm Spectral Monchrometer Grating
Channel 2 SDM640, SDM560 2nm Spectral Monchrometer Grating
Channel 3 SDM640 BA560-620, BA560-660, BA655-755

Zeiss Cell Observer SD Spinning Disk Confocal

zeiss SD

Location: 1-160 Jackson Hall

Spinning disk confocal fluorescence microscope for imaging live samples at fast speed. Definite focus, time-lapse, multi-well and tiling of images. DIC and phase contrast capable. Full environmental chamber for temperature and atmosphere control. Three laser lines available (405, 488, 561 nm). 

Learn more about the Zeiss Cell Observer SD Spinning Disk Confocal

Motorized Zeiss Axio Observer Z1 inverted scope with temperature controlled enclosure, CO2, Definite Focus, multi-channel acquisition, piezo x/y/z stage.

Methods Available:

  • Fluorescence
  • Brightfield
  • Phase contrast 
  • DIC
  • Time Lapse
  • Live Cell with 37 degree C / CO2 enclosure

Automated Capabilities:

  • Time-lapse
  • Stitching
  • Multi-channel fluorescence
  • Co-localization
  • Piezo Z-stacks

Objectives:

Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
10x 0.3 EC Plan-NEOFLUAR Ph1 ∞/- 3.2 Air
20x 0.75 Plan-ApoCHROMAT Phase 2, DIC II ∞/0.17 0.61 Air
40x 0.95 EC Plan-APOCHROMAT KORR ∞/0.13-0.21   Air
63x 1.4 EC Plan-ApoCHROMAT   ∞/0.17 0.19 Oil Immersion
63x 1.2 C-ApoCHROMAT KORR UV-VIS-IR, Correction Collar ∞/0.17 0.28 Water Immersion
100x 1.4 Plan-ApoCHROMAT DIC ∞/0.17 0.11 Oil Immersion
Lasers: 

405 nm, 50 mW
488 nm, 100 mW
561 nm, 30 mW

Detector: 
  • Photometrics QuantEM 512SC - High Sensitivity 16 bit, 512 x 512 grayscale CCD

Snyder Hall

Nikon A1 Spectral Confocal

Nikon A1

Location: 35D Snyder Hall

The Nikon A1si is a confocal system equipped with a point-scan scan head, 5 standard PMT detectors and a 32-channel PMT spectral detector. The A1si's highest resolution mode is up to 4096 x 4096 pixels. The system is mounted on a Nikon Eclipse Ti2 inverted, fluorescence microscope with DIC optics. NIS Elements imaging software is used to control acquisition and analysis.

Learn More about the Nikon A1 Spectral Confocal

Advantages of multispectral imaging and unmixing has been shown in hundreds of publications to reduce autofluorescence by as much as 100-fold, both for microscopy and FFPE tissues and in small animal and plant fluorescence imaging. 

A few example citations are:

Tam, et al "Improved in vivo whole-animal detection limits of green fluorescent protein-expressing tumor lines by spectral fluorescence imaging" Molecular Imaging, 6, 2007, 269.

Mansfield, et al "Visualization of microscopy-based spectral imaging data from multi-label tissue sections" Current Protocols in Molecular Biology, 2008 Oct;Chapter 14:Unit 14.19. doi: 10.1002/0471142727.mb1419s84.

Levenson, et al "Multispectral imaging in biology and medicine: slices of life." Cytometry A. 2006 Aug 1;69(8):748-58.

 

Objectives:
Magnification NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
4x 0.2 Plan Apo -- ∞/-- 20 Air
10x 0.3 Plan Apo -- ∞/0.17 4 Air
20x 0.75 Plan Apo DIC N2 ∞/0.17 1 Air
60x 1.2 Plan Apo DIC N2, Correction collar ∞/0.15-0.18 0.27 Water Immersion
60x 1.4 Plan Apo DIC H ∞/0.17 0.21 Oil

Fiilters:

Filter Wavelengths
Excitation Filter (nm) Dichoric Mirror (nm) Emission Filter (nm)
DAPI DS350/50 BS20/80 460/50
FITC D480/30 DM405/488 D535/40
TRITC D540/25 408/488/543 D600/50
Cy5 D640/20 DM458/515 D685/70
Channels Dichoric Mirror (nm) Barrier Filters (nm)
Channel 1 SDM560,SDM510,SDM490 2nm Spectral Monchrometer Grating
Channel 2 SDM640,SDM560 2nm Spectral Monchrometer Grating
Channel 3 SDM640 BA560-620,BA5605-660,BA655-755
Lasers: 
  • 405 nm diode at 38mW
  • Argon multiline laser 457/477/488/514nm at 40mW
  • 561 nm diode at 40mW
  • 638 nm diode at 10mW
Detectors: 

4 PMTs, Transmitted light detector and 32-channel spectral detector.

Nikon AZ100 C1si Spectral Confocal Macroscope

az100 c1si

Location: 23B Snyder Hall

The AZ100 Multizoom Macroscope with the C1si Spectral Confocal attachment, make the ultimate platform for in vivo imaging. Ideal for plant biology, developmental biology, cell biology, stem cell and tissue research, the AZ-C1si allows the researcher to view large specimens in confocal mode. The AZ-C1si can capture fields of view up to 1cm and permits deeper confocal imaging than conventional microscopes thanks to its large working distance objectives. Whole organisms can be monitored and documented over time (for example, embryos) offering a wealth of continuous information on development or the organism’s response to experimental variables. Both conventional PMTs and Spectral detectors are installed on this instrument.

Learn More about the Nikon AZ100 C1si Spectral Confocal Macroscope

The system is equipped with 1x, 4x and 5x objectives with variable optical zoom.  There are 405, 457, 476, 488, 514, 561, 637 nm lasers for image, co-localization, FRAP, FRET and spectral studies.

The system is run using Nikon Elements Software.

Advantages of multispectral imaging and unmixing has been shown in hundreds of publications to reduce autofluorescence by as much as 100-fold, both for microscopy and FFPE tissues and in small animal and plant fluorescence imaging. 

References:

Tam, et al "Improved in vivo whole-animal detection limits of green fluorescent protein-expressing tumor lines by spectral fluorescence imaging" Molecular Imaging, 6, 2007, 269.

Mansfield, et al "Visualization of microscopy-based spectral imaging data from multi-label tissue sections" Current Protocols in Molecular Biology, 2008 Oct;Chapter 14:Unit 14.19. doi: 10.1002/0471142727.mb1419s84.

Levenson, et al "Multispectral imaging in biology and medicine: slices of life." Cytometry A. 2006 Aug 1;69(8):748-58.

Objectives:

Magnification  NA Correction Optic Properties Tube Length/Cover Glass Thickness Working Distance (mm) Immersion Medium
1x 0.1 AZ Plan Apo     35 mm Air
4x 0.4 AZ Plan Apo     20 mm Air
5x 0.5 AZ Plan Fluor     15 mm Air

Filters:

Filter Wavelengths
Excitation Filter (nm) Dichoric Mirror (nm) Emission Filter (nm)
DAPI D350/50 BS20/80 460/50
GFP D480/30 DM405/488 D535/40
TRITC D540/25 405/488/543 D605/55
Cy5 D640/20 DM458/515 D680/30
Channels Dichoric Mirror (nm) Barrier Filters (nm)
Channel 1 SDM560, SDM510, SDM490 2nm Spectral Monchrometer Grating
Channel 2 SDM640, SDM560 2nm Spectral Monchrometer Grating
Channel 3 SDM640 BA560-620, BA560-660, BA655-755

Lasers:

405
457
488
514
567
637