Zeiss Axio Imager widefield fluorescence microscope

Now upgraded as this system

Duke LMCF Zeiss Axio Imager microscope Capabilities:
  • Widefield fluorescence imaging (eg Dapi, CFP, GFP, YFP, Alexa 488/568/633)
  • Transmitted light images - brightfield, DIC and polarized light
  • Colour camera for IHC (eg H&E staining)
  • Ideal for thin fixed specimens (oil objectives)
  • Z-stacks

Location: Room C171 LSRC (613-8217)

User manual: pdf | html | quick_guide.pdf

 

Detailed specifications

Upright AxioImager.A1 microscope stand

Mercury arc lamp: Zeiss HBO100 power supply & lamp housing

Filter cube sets (turret):

  1. Multipass dichroic (Semrock Pinkel r381-392/t420-460, r475-495/t510-531, r547-572/t589-623, r643-656/t677-722)
  2. DAPI, Hoechst (Chroma 31000v2 D350/50x, 400DCLP, D460-50)
  3. CY5 Zeiss Filter Set 50 (488050-9901-000), BP 640/30, FT 660, BP 690/50
  4. Green, GFP, FITC, Alexa488 (Zeiss Filter Set 38 HE 489038-9901-000, BP 470/40, FT 495, BP 525/50)
  5. YFP (Chroma 41028 HQ500/20, Q515LP, HQ535/30)
  6. Red (Chroma 41004 Texas Red, Alexa 568, Alexa 594 HQ560/55, Q595LP, HQ645/75)

CFP cube is available (Chroma 31044v2 D436/20, 455DCLP, D480/40)

Chroma 41001 FITC/Bodipy/Fluo3/DiO (HQ480/40, Q505LP, HQ535/50) available (GFP very similar)

Excitation filters (filterwheel A):

Use with Semrock quadpass dichroic

  1. Open (no filter)
  2. Closed
  3. 485/20 (FITC, Alexa 488)
  4. 560/25 (TRITC, Alexa 568)
  5. 387/11 (DAPI)
  6. 650/13 (Cy5, Alexa 647)
  7. 425/40

Neutral density filters (filterwheel B):

  1. Closed
  2. Open (100% Transmission)
  3. 80% T
  4. 60% T
  5. 40% T
  6. 20% T
  7. 10% T

LUDL emission filterwheel:

  1. Open
  2. 684/24
  3. 624/40
  4. 531/22
  5. 447/60
  6. Analyzer

(Semrock Pinkel multipass emitter was 440/40, 520/21, 606/34, 699/45)

Objectives:
  • 2.5x/0.075 Dry EC Plan NeoFluar 440310-9903 WD 9.5 mm
  • 5x/0.16 440320: EC Plan-Neofluar, NA: 0.16, air, WD: 18.5mm
  • 10x/0.30 440330: EC Plan-Neofluar, NA: 0.30, air, WD: 5.2mm
  • 20x/0.50 DIC 440340: EC Plan-NeoFluar, NA: 0.50, air, WD: 2.0mm, coverglass: 0.17mm
  • 40x/0.75 DIC 440450: EC Plan-NeoFluar, NA: 0.75, air, WD: 0.71mm, coverglass: 0.17mm
  • 63x/1.4 oil DIC 420760-03: Plan-Apochromat, NA: 1.40, oil, WD: 0.19mm, coverglass: 0.17mm, thread type: M27x0.75
  • 100x/1.4 oil DIC 420780-02: Plan-Apochromat, NA: 1.40, oil, WD: 0.17mm, coverglass: 0.17mm, thread type: M0.8X1/36
  • 5x px = 1.2407 micron for Orca, 1.0406 micron for color camera
  • 10x px = 0.6261 micron for Orca, 0.5223 micron for color camera
  • 20x px = 0.3120 micron for Orca, 0.2622 micron for color camera
  • 40x px = 0.1520 micron for Orca, 0.1296 micron for color camera
  • 63x px = 0.0976 micron for Orca, 0.0807 micron for color camera
  • 100x px = 0.0593 micron for Orca, 0.0524 micron for color camera

 

Cameras:

  • Hamamatsu Orca ER monochrome cooled-CCD camera with IEEE (firewire)
  • QImaging MicroPublisher 5.0 MP colour camera

Computer:

  • Dell Optiplex 745 Intel Core 2 Duo processor E6600 2.40GHz, 4 GB RAM, Windows XP Pro 32 bit
  • MetaMorph 7.6.5, system ID 3551