(JULY 1998)

Start up

1. Turn on the spec and monitor and let warm-up for at least 30 minutes in level of light the samples will be run.
2. While waiting, set up lake sets and centrifuge samples.
3. After the warm-up, re-run the baseline by selecting 1, Uvikon start, on the main menu page and highlighting NEW SYSTEM BASELINE. This will take about 5 minutes to run.
4. Check the baseline by running blank methanol. Highlight USER METHODS then highlight LTER003. On the first parameter page check that the sample number is at 1 and autosave is off. Place cells filled with optima methanol in both the reference position and the sample position and press MEASURE. It should be a straight line at + 0.005. Refer to LTER chlorophyll protocol if not.

Registering Cuvettes

5. Line up the cuvettes in order to begin registering
6. Change the number of samples run and turn autosave on
7. Place optima methanol in the first cuvette so that it may be transferred from cuvette to cuvette to minimize the use of methanol. Place cuvettes in the sample holder.
8. Begin running cuvettes in the order they are numbered. Keep track of how many sets of samples are run throughout the spec run to later go back and rename.

Running Samples

7. Align samples with numbered cuvettes in order of lake set.
8. Transfer sample from centrifuge tube to cuvettes using a disposal pipette. Rinsing in between sets is not recommended unless the color change is noticeable. Do not disturb filter slurry and make sure none of the filter is transferred.
9. Place sample in cuvette holder and press measure. Keep track of the order of the sample sets. Each lake has its own file. Refrigerate any remaining sample in a dark box as soon as possible.

Acidifying and neutralizing

10. Place the 1.0N HCl syringe in repipetter. Set repipetter to dial 1, for acid addition of 25 uL. Use a glass stir rod to mix the sample well and then wipe the glass rod with a paper towel in between sets. Wait 3 minutes after last acid addition for the reaction to take place.
11. Place the 1.0N NaOH syringe in repipetter. Set repipetter to dial 4, for base addition of 40 uL. Use a glass stir rod to mix the sample well and then wipe the glass rod with a paper towel in between sets. Wait 1 minute after last base addition for neutralization.
12. Run post-acid samples as normal remembering to keep track of lake set and file order.
13. When finished empty cuvettes and rinse with bulk methanol and allow to drain and dry.

Renaming and Transferring Files

14. Select METHODS to return to the main screen.
15. Highlight DISK AND FILE UTILITIES and select 9 to EXIT DOS.
16. At prompt type "cd windows" ENTER and then type "win" ENTER.
17. Open FILE MANAGER and then open the WORK folder.
18. Sort files by name and highlight the most recent file ran.
19. Hold down ALT-F then press N to rename the files.
20. Rename samples one at a time following the format:

a. Pre-acid YR,JULIANDAY,LAKE (excluding commas)

b. Post-acid YR,JULIANDAY,LAKE,"A" (excluding commas and quotes)

Example: pre 98122BM post 98122BMA

21. Highlight all renamed files and rename as " *.scn". This will add .scn to the end of the files.
22. Exit windows and at dos prompt type "9310".
23. Select 3 for File Format Conversion, then select 1 for ASCII Format and then 4 for Wavelength Scan Data. Highlight all files by pressing ENTER. Press EXECUTE.
24. Exit by pressing 7, then 3, then 9 to exit to dos.
25. At C:\> type "novell" and log in as normal.
26. Type "nm" to enter network. Select windows <A> then select local windows <B>.
27. Open file manager and also open O:/lter/phys/chloro.
28. Next open the C directory, sort by type, and highlight all .asc files. Drag highlighted files to the O directory. Delete .asc files.
29. Open C:/lter_chloro/1998 or most recent year. Return to C:/work and highlight .scn files. Drag highlighted files to 1998 folder.
30. Exit windows, ESC to menu and press K to log out of network.
31. At prompt type "9310" and turn off spec.