Spectrophotometers:

• There is one to the left of the bench.

Ø If it is not already turned on, turn it on with the switch on the bottom right of the machine (fig. 1).
Before you put the cuvettes in, press “auto zero” on the control panel of the spectrophotometer
making sure that the absorbance screen on the control panel shows zero (sometimes you need to
press the button twice).
Ø Put the cuvette containing the reference solution in the rear holder and the sample in the front one
(fig. 2). You have to dilute the sample to concentrations of about 0.1-0.5 mg/ml using the denaturing
UV cocktail you can find on the bench.
Ø Before you start make sure no other measurement windows are open (if there are and none seems to
use the spectrophotometer close them).
Ø To change the settings, the type of measurement etc. go to MethodsàNew Method nad choose the
desired type of method (fig. 3).
Ø You can also load a user method such as Bradford etc (fig. 4).
Ø By pressing the button Edit at the bottom of the screen you can change various parameters of the
method such as wavelength etc (fig. 5). The wavelength can also be changed from the control panel.
Ø Then press the “M” measure button on the upper part of the screen (fig. 6, 7). You can also see the
absorbance (if using fixed wavelength) on the control panel under the “Abs” tag. The measurements
can also be saved or printed (press File on top of the screen).
Ø If you want to stop the measurements (e.g. in case of a wavelength scan) press the stop button next
to the measure button (fig. 8).
§ This is the only spectrophotometer that has temperature control. If you want to use the temperature
control unit it can be found on the left of the spectrophotometer.
ü Turn it on.
ü If there is no water left then fill it with demineralised water.
ü Also the flow switch should be turned on (remember to turn it off when done).

• The other one is in the smaller room (fig. 9).

Ø By using the “mode” button and the arrow keys you can navigate through different modes like
fixed wavelength measurements, wavelength scan etc.
Ø With the “methods” button you can select different methods of measurements such as Bradford.
There a lot of parameters that can be changed in this one, it can also be used for kinetics.
Ø For example for a fixed wavelength press “mode” then “enter” then choose the submenu
“absorbance” using the arrow keys and press the “wave” button to set the wavelength using the
numerical keypad. Press “enter” to save your changes.
Ø This one has eight (8) sample holders (usually the first one (blue) is used for the reference). To set
to zero the absorbance for the reference solution
press “set ref” (the spectrophotometer moves
automatically to the next sample holder). To select
a sample holder just press the desired number on
the control panel. The absorbance can be seen on
the screen and also printed using the “print” button
(press “stop” when you are done before you cut the
paper).

Rear
holder

Front
holder
Control
panel
1
2
3

4
5

6
7

8
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