Chapter 9: Procedures and Technical References
Adding/Replacing Filters
•Physical Addition/Replacement of Filters
–Excitation Module
1. In order to avoid having to realign the filters encoders, do not shut down
power to the Nanomotion chassis.
2. Remove fiber optic cable from condenser lens on Excitation Module
3. Place cable in a location where it will not be bumped
4. Unplug the EX shutter cable
5. Remove six screws module from the motor side (two top, two middle, two
bottom. Take care when tilting module to prevent screws from falling
anywhere but the table or your hand.
6. Set the module in a position where the filter may be accessed. Take care to
prevent the motor cable from being pinched or otherwise damaged.
7. Cotton gloves are recommended for handing filters. Fingerprints may not be
removed once they are on the coated portion of the glass.
8. Loosen, but do not remove, the three screws in the filter holder for ND
filters. For EM and EX filter spread the hold down springs. (Some filters will
require removing one of these screws to allow more height.)
9. Rotate and remove the holder
10. Remove the block or old filter.
11. Place the new filter such that the arrow on the side of the filter is pointing
toward you. (Both Chroma and Omega filters.)
12. Note the location if adding new filters.
13. Slide the holder back on to the screws and set well For EX and EM filters
make sure springs seat firmly over sides of filter.
14. Tighten the screws such that the metal is slightly bowed but not enough so it
starts sliding back away from the screws (ND only).
15. Place the wheel portion of the module back on the lamp portion, replace, and
tighten the six screws.
16. Place shutter cable back in the EX motor board.
17. Reattach the fiber optic cable.
–Emission Module
1. Remove Emission Module cover.
2. Mark module position on table with tape or a marker. This will assist in
aligning the camera.
3. Remove the screws holding the module to the table and pull module away
from scope.
4. Remove four screws holding filter cover on module. TAKE CARE TO
PREVENT TWO TOP SCREWS FROM HITTING ANY OF THE
FILTERS!
5. Cotton gloves are recommended for handing filters. Fingerprints may not be
removed once they are on the coated portion of the glass.
6. Spread the filter hold down springs.
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Chapter 9: Procedures and Technical References
7. Remove the block or old filter.
8. Place the new filter such that the arrow on the side of the filter is pointing
toward you for Chroma filters and away from you for Omega filters.
9. Note the location if adding new filters.
10. Make sure springs are over filter edges and hold filter firmly in place.
11. Replace the filter cover and tighten four screws.
12. Align Camera
a. Move eyepiece lens to open (O.)
b. Slide the emission module up to the microscope and start but do not
tighten two screws and washers in hold down bar (or four screws in
legs for older versions.)
c. Place the mirrored slide on the stage.
d. Set the microscope to reflective mode (Excitation filter set to RD-FTPE, emission filter set to FITC) and set the ND to 3.0.
e. Focus on the scratches on the slide.
f. Close the field aperture.
g. Move the viewing knob to SP (side port.)
h. Select size 512x512 and bin 2 in Resolve3D.
i. Acquire series from Resolve3D using an exposure time that does not
saturate the camera.
j. Center the hexagonal image in the display window by moving the
camera side to side for up and down in the SGI window.
k. If the side to side in the SGI window is not aligned slightly loosen four
screws on back of the EM module face plate and raise or lower the
plate with the upper vertical screws. Retighten four screws when
complete.
l. Bolt the camera to the table.
m. Stop Series in Resolve3D.
•Modify the Software to Reflect New Filters
Prior to making any changes to any software, make a copy of the file
as a back up.
–Dvpc408.ini or ic510.ini
1. Open the appropriate file, in ic5XX.ini, with Notepad on the PC.
2. Scroll down and find the following sections of code. TAKE CARE TO
MODIFY ONLY THE NECESSARY PORTIONS. Other additions may
crash the code.
[Excitation Filter Wheel]
…
Number of Filters = 10
Filter Names = BLANK1 DAPI FITC RD-TR-PE CY-5 BLOCK6 BLOCK7 BLOCK8 BLOCK9
BLOCK10
Filter Values 1= 0. 360. 490. 555. 640. 0. 0. 0. 0. 0.
Filter Values 2= 0. 40. 20. 28.
20. 0. 0. 0. 0. 0.
Filter Pos= 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00 8.00 9.00
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Chapter 9: Procedures and Technical References
[Emission Filter Wheel]
…
Number of Filters = 8
Filter Names= BLANK1 DAPI FITC RD-TR-PE CY-5 BLOCK6 BLANK7 BLANK8
Filter Values 1= 0. 457. 528. 617. 685. 0. 0. 0.
Filter Values 2= 0. 50. 38. 73. 40. 0. 0. 0.
Filter Pos= 0.00 1.00 2.00 3.00 4.00 5.00 6.00 7.00
3. Add the filters with the exact location of actual placement in the wheel.
4. Add both the wavelength and band pass in the location relating to the
appropriate filter.
5. Use only the first position (0.00, BLANK1) when all other positions have
been filled. These positions are used for wheel alignment and should be kept
open if possible.
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AppliedPrecision