SOP of NEPHELOMETER

NEPHELOMETER
STANDARD OPERATING PROCEDURE

1. Turn on the instrument and the computer.

2. Start the Omega software. Login with your password or just click RUN to login as USER.

3. To measure a microplate, insert the microplate by the command Plate In.

4. To create a new Test protocol or to edit an existing one:

Click the Manage Protocols button.
Double click the protocol name to edit an existing protocol or click New to create a new protocol. Choose the Measurement Method and Reading Mode.

5. Inside the protocol definition window:

Enter a protocol name.
Choose the microplate being used.
Select between Rapid or Precise measurement, depending on your assay.
Plate Mode Kinetics: type in the Number of Cycles (how many times the reader will cycle through the plate).
Well Mode Kinetics: type in the Number of Intervals (how many times the reader will read the well).
Choose the Excitation and Emission Filters to be used.
Select the Layout sheet. Enter the position of samples, blanks and standards.
If standard and / or reagent dispenser are used, type in the values in the Concentratin/Volumes/Shaking window.
Click the Check Timing button. This gives you the smallest possible cycle time (plate mode) or interval time (well mode). A longer time can be achieved by typing in a higher value in the Basic Parameters sheet.

6.Click the Start Measurement button.

7. To see the measurement results during a reading. Click the Current State button.

8.To perform data calculations using the Mars Data Analysis software.

9.Double click on the Test name of the test run to be analyzed.

10.Analyze the measured data:

Select the data to be displayed in the working area with the navigation tree (Data Node) on the left side of the main window.
Use the standard calculation wizard to perform a quick curve fit calculation; or use the calculation menus to define what is to be calculated and to be displayed.
To see the Standard curve, open the Standard Curve page. The calculated unknowns are displayed in the Microplate View and the Table View.
For kinetic measurement (more than one measured cycle or interval), choose the ranges of interest (calculation Start and Stop) and the data values from these ranges can be evaluated using a kinetic calculation.