Capabilities

• Widefield imaging of living and fixed samples
• Optimized deconvolution
• Solid-state illumination, fast excitation switching
• Temperature, humidity, CO2 control
• Hardware-based autofocus
• High-precision XYZ stage
• Dual cameras - high-resolution and high-sensitivity
• 405 nm and 488 nm lasers for photoactivation and photo-bleaching experiments
• Optical axis integration/z-sweep acquisition
• Kohler and critical illumination

Detailed Specifications

The EM camera is currently removed from this system.

Link to API website

Microscope:

• Olympus IX-71 base
• API illuminator with Kohler and critical epi illumination
• LWD DIC condensor with LED illumination
• Ultimate focus - hardware based focus stability
• High precision XYZ stage with inserts for slides and 35 mm dishes
• Opaque Weather Station incubation system with regulated temperature, CO2 and humidity

InsightSSI 7 channel solid state illumination source with fast switch times:

• Fiber coupled for homogenous scrambled light illumination
• Monitor diode
• DAPI 390/18
• CFP 438/24
• GFP/FITC 475/28
• YFP 513/17
• TRITC 542/27
• mCherry/AF594 575/25
• Cy5 632/22

Fluorescence attenuator:

• Open (100% Transmission)
• 50% T
• 32% T
• 10% T
• 5% T
• 2% T

Emission filterwheel 10 position:

• Blank (for high speed)
• CFP 470/24
• YFP 559/38
• mCherry/AF594 632/60
• GFP/FITC 525/50
• DAPI 435/48
• TRITC 594/45
• CY5 676/34
• Analyzer (pol)

Eyepiece filter wheel 7 position (manual and encoded)

• POL Analyzer
• DAPI
• TRITC
• GFP/FITC
• mCherry/AF594
• CFP
• YFP

Filter cubes/dichroics:

1. Polychroic BGRD,FR (standard quad) DAPI,FITC,TRITC,CY5
2. Polychroic BGmCherry,FR (alexaquad) DAPI,FITC,AF594,CY5
3. CFP/YFP/mCherry
4. GFP/mCherry (Chroma)
5. Dual em pass CFP/YFP for high speed
6. Dual em pass GFP/mcherry for high speed

Objectives:

• 10x/0.40 UPLAPO dry 1-U2B823 WD 3.1 mm ∞/0.17/FN26.5, UIS2
• 20x/0.70 UPLAPO dry 1-U2B825 WD 0.65 mm DIC ∞/0.17/FN26.5, UIS2
• 40x/0.65-1.35 oil UAPO40XOI3/340 1-UB768R WD 100 micron DIC ∞/0.17/FN22, UIS2, BFP1
• 60x/1.42 oil PLAPON60XO WD 150 micron DIC ∞/0.17/FN26.5, UIS2, BFP1
• 60x/1.20 water UPLAPO60XW/PSF 1-UB891 DIC WDWD 250 micron ∞/0.17/FN26.5, UIS2
• 100x/1.40 oil UPLSAPO100X0 1-U2B836 WD 120 micron DIC ∞/0.17/FN26.5, UIS2

Stage

• 50 mm travel in Y, 25 mm in X, 1 mm in Z

Cameras:

• Coolsnap HQ2 high resolution CCD camera
• Evolve back-thinned EM-CCD camera (512*512)

Lasers for photoactivation and photobleaching:

• 488 nm 50 mW
• 405 nm 50 mW

Computer:

• Dual Xeon processors, 16 GB RAM, 2 TB RAID data array and 250 GB boot drive, 30" monitor
• Linux (Centos 6.5) 64-bit
• SoftWoRx 6.1, with system-level queuing
• Similar system for offline deconvolution

This system was purchased with funding from the NIH/NCRR Shared Instrumentation Grant 1S10RR027528-01