I. Make sure the instrument is turned on.

There is a green rocker switch on the back right corner of the top of the instrument. It is on when the front part of the switch is pressed down.

II. Double click on the Lambda Bio 40 icon on the desktop to start the software.

When the software starts, a big window opens. Inside that window, there is a smaller Methods window that opens, and three windows along the bottom - Data Region, Results Wind(ow), and Graph1. There is a line at the bottom of the window that reports the instrument status.

III. Choose a method either from the methods window or by clicking one of the buttons above.

 You will see that the Methods window has a series of five tabs along the bottom. They sort the methods into Scan, Time Domain (Td), Wavelength Programs (Wp), Concentration (Conc) and Others. 

First choose a tab, then choose a method. If you click once on a method name, a brief description of the method appears on the line under FileInfo. Each category of method has a default method that corresponds to the button choices at the top of the screen:

Scan Td Wp Conc

If you click on one of the buttons, the corresponding default method will open.

4. Make any needed changes in the chosen method.

The method tab allows you to adjust parameters directly related to the specific method. The Inst tab controls more basic instrument function such as which lamps are turned on, the scan speed, slit width (narrower slit = higher resolution), and smoothing functions or response time. It also allows you choice of %T or A as the initial display mode. This may be changed after data acquisition as well. The Sample tab controls naming of the data files, number of samples to be used, and "factor" - typically just left at "factor."

V. Do the experiment.

Open the cover on the top right side of the spectrometer and insert your samples as instructed by the software. Usually you will need to put a blank sample in the rear cuvette holder and your sample to be measured in the front holder.
  A. Wavelength Scans 

It is a good idea to perform an autozero before actually collecting data. In this case, you click on the Autozero button and, when prompted, insert a blank sample in both the front and rear holders. When the Autozero is complete, the Start button will be green. Click on the Start button. You will be prompted to insert your sample (in the front holder). Then click Okay and the scan will start. When it is done, the Start button turns green again. Your results appear in the Graph1 window. You can use the buttons at the top to start the report builder, set the XY scale, set to previous XY scale, show a "radar" window for navigating a zoomed region, rescale left-right, rescale vertically, find peaks, insert a cursor (which you can drag to read X & Y values), and add text. You can zoom a region by left clicking on the spectrum and dragging to make a green box. Double click in the green box to zoom it. If you want to overlay results from previous scans, look in the Data Region window and click on the scan to be added to the display. If you want to save your results choose File -> Save As. A new window will pop up. You can navigate to the directory you want ([..] means go up one level) and then save. There are four possible formats - binary, Data manager, ascii, and JCAMP. All types are recognized by the Perkin Elmer software (you can reopen old spectra), but JCAMP or ascii are the most useful to transfer to other programs such as Excel.
  B. Time Domain 

The default time domain scan monitors the signal level at one wavelength as a function of time. To run the experiment, you need to choose the desired wavelength and enter it on the Timed page. Enter other parameters as needed, checking all the tabs. Click Start. You will be asked to insert the blank, then prompted for the sample. The data appears as a graph of signal level versus time.
  C. Wavelength Program 

The default wavelength program allows you to program up to eight individual wavelengths for repeated measurements. You enter the number of wavelengths, the number of cycles (repeats of the readings) and the time interval between cycles (cycle time) on the Wavep tab. Note that the minimum cycle time will be determined by the number of wavelengths you measure. If the cycle time you enter is shorter than the time the instrument needs to execute the cycle, the next cycle just starts as soon as possible. The Inst and Sample tabs control the other instrument parameters. The first request upon clicking the Start button is to insert a blank for baseline correction. Results are written in the Results Window that automatically opens.
  D. Concentration  The default concentration method allows you to determine concentrations of a series of solutions by a variety of methods including single wavelength, single wavelength + peak, derivative, derivative + peak, area, and ratio. There are also a variety of baseline correction methods and curve fitting options. The user is advised to look at the Perkin Elmer tutorials for more information.

VI. Analyze and Save Your Results

Depending on the type of experiment you have performed and which window is active (i.e. Graph1 or Results) you will see different buttons available to help you process your data. For example, if you did a Wavelength Scan, your results will appear in the Graph1 window, and you will see buttons like this:

These buttons allow you to start the report generator, resize the axes, restore the axes to the previous values, insert a "radar' window to show you where the zoomed region is relative to the whole spectrum, resize the horizontal or vertical axes to full scale, find the peaks, insert a vertical cursor that allows you to read the X & Y values of your spectrum, or insert text. In addition to using these buttons, you can use the Data Handling menu to access other processing capabilities. You can find the peaks (Peak), list the spectral intensities every 0.5 nm (List), or switch back and forth between absorbance and % Transmission (A <>T). You can also use the Data Calculator menu to perform different math operations on one or more data sets. These operations include baseline smoothing, finding peak areas, interpolation, normalization, taking the derivative of spectra, and arithmetic functions such as adding two spectra.

If you want to save your results choose File -> Save As. A new window will pop up. You can navigate to the directory you want ([..] means go up one level) and then save. There are four possible formats - binary, Data manager, ascii and JCAMP. All four types are recognized by the Perkin Elmer software (you can reopen old spectra), but JCAMP or ascii is the most useful to transfer to other programs such as Excel.