Nikon Eclipse TE2000 Spinning Disc
Nikon Eclipse TE2000 Spinning Disc
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inverted perfect focus microscope
optimized for live cell imaging and time lapse movies
Equipped for DIC, Ludl Piezo Z-stage for 3-D imaging
temperature controlled chamber
widefield cubes for GFP, YFP, TxRed, CFP, DAPI
Cascade 512B and Coolsnap HQ CCD detectors offer
simultaneous data acquisition
• 488 and 568 nm Ar lasers
• Visitech QLC100 confocal head.
Turning on and Setting up the Microscope
1. Turn on Power to the AOTF Controller
2. Turn on Power to the Laser Power Supply
*DO NOT TOUCH THE KEY!
3. Turn on Power to the Laser Supply Remote
Controller:
a: Flip switch to “operate”
b: Turn key past “on” and let go
c: Laser should go to “100” or more
d: Let warm up 30 min before using
e: Flip switch to “standby” if not
using the lasers for awhile or when
you leave
f: Laser Power can be displayed in
mW or in AMPs depending on which
position the toggle is flipped
g: Laser Power can be changed by
toggling the switch up (increasing
power) or down (decreasing power)
4. Open AOTF Controller Software
- Adjust % Power in software if
needed
- Usually start with 100%
- Must do this before doing next
steps
5. Turn on Black APS Power Supply
6. Turn on Temperature Contoller:
- Turn dial to “set” option and use
dial to set control temperature.
- Keep at 39oC so that the center of
your dish is 37oC.
- Turn dial to T1 to see actual temp.
7. Turn on Ludl Shutter Control:
- Widefield/DIC is Halogen bulb
- Fluorescence is mercury bulb
8. Turn on Xcite Mercury Lamp
9. Turn on Halogen Lamp
*This lamp is under the air table.
10. Turn on confocal head key
*Both lights should be on.
11. Turn on the Perfect Focus Box:
- Push green button to “engage”.
- If green light flashes, it is not
working properly.
- Be sure to “disengage” PF by
pushing green button before
removing your dish of cells from
the stage.
12. Turn on Power to the Microscope:
- Back left bottom of scope
where red tape is located.
13. Turn on Camera(s) before opening IPLAB:
- Cascade is for Confocal
- CoolSnap is for Widefield/DIC/FL
14. Open up IPLAB Software:
*You should always get an error
message. It is ok.
15. Tell software which camera you’re using:
- Select “Camera” option
- Select “Select Camera”
- Click “OK” by camera
16. Setup camera settings:
- Select “Camera” option
- Select “Get Camera Info”
- Can adjust settings
- Gain Index = 3
- Speed Table = 1
- Mult Gain = 3700
17. Adjust Magnification Dial:
- Located on bottom right of
scope.
- “1x” means no additional
magnification.
18. Plug in heater controller to stage dish:
- Plug temperature probe into
small hole on side of setup.
- Attach blue clips to each side.
- Use the gold ring insert for most
MatTek dishes.
*Other ring inserts available.
Turn off Microscope
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Turn lasers to zero in AOTF Controller Software.
Turn off Power to Perfect Focus box.
Flip lasers to “standby”.
Turn laser key to “off”.
*Listen for fan to get quiet before turning anything else off.
Remove sample and clean-up. Liquid waste is nearby and
should be emptied if needed.
Leave the stage dish in place.
Turn off the rest of the equipment once hear fan power
decrease.
*IPLAB must be closed before camera is turned off.
Put cover on microscope.
Sign log sheet.