IMB205

 Yuan’s lab protocol for Aviv Model 400 CD spectrophotometer

1.) Ask the CD manager for appropriate cuvettes and cuvette spacer.

2.) Open the nitrogen valve with the green screw top and check the pressure gauge indicating at 20 psi the flow rate should be between 15-20 SCFH, waiting for 20 minutes to purge the system.

3.) Switch on the water bath at right hand side of the CD

4.) Switch on the switch ”lamp power and cooling system” on power box, the led light ”Control power On” will be on.

5.) Once the “lamp ready” is on use the “Ignite Lamp” to ignite the lamp. “Lamp On” lit indicate the light source is on.

6.) Write down the hours from the display into the users-book.

7.) Start “CPU & instruments”. Just press enter when the computer asks for a password.

8.) Start computer software from the desktop and load the configuration file at the menu “File”.

9.) Set the data path under the Menu “Displays ->Data Browser ->Default Dataset Path” to c:/Documents and Settings/AVIV/My Documents/hanna205/your personal folder/file names.

10.) Open “Configure Experiment” from the menu, term “Description” and “Name of the data”. When changing temperature, you have to push enter after setting the temperature. With “Experiment Configuration” the other settings like wavelength can be adjusted. Click on “Exit/Save Configuration” to leave this window.

11.) Put the cuvette into the slot, close the box and push “Run Experiment”.

12.) Save the experiment in data browser and hard drive.

13.) Go on with “Axis Definitions” and choose “Data Collection Average”. Mark all shots and click “Average Selected Traces”. Then save average trace and close the window.

14.) To start a new experiment go to “Configure Experiment” and change name of the file and the conditions. If changing the temperature, wait till the new temp is reached (about 20 sec. for 5 degrees) and wait 3 minutes for equilibration.

15.) When all experiments are done go to “Displays/Data Browsers”. Click the Wavelength folder and save your files as .txt files on your USB drive.

16.) Cuvette clean-up: (i) rinse the cuvette with mQ water, (ii) pipette appropriate premixed solvent: sulfuric acid/ EtOH/water into the cuvette and incubate overnight for effective clean-up (iii) tip off the cleaning solvent and rinse with mQ water & (iv) dry it up with air pump.