biochrom WPA Lightwave 3 User manual

W
Lightwave 3 & Lightwave 3+
Spectrophotometers
Users Manual
5061-084-REV-1

LIGHTWAVE 3 & 3+ SPECTROPHOTOMETERS USER’S MANUAL
2
TABLE OF CONTENTS
5061-058-REV-11
WARRANTY AND REPAIR 4
Warranty Policy 5
SAFETY INFORMATION 6
Hazards and Warnings 6
INTRODUCTION 8
The Biochrom WPA Spectrophotometers 8
INSTALLATION 9
Unpacking 9
Positioning 9
Installing 9
INSTRUMENT OVERVIEW 10
Scope 10
Spectrophotometer Principle and Intended Use 10
Hardware 10
Technical Specications 11
Display and Keypad 11
Instrument Connections 11
PVC PC Software 12
Biochrom Resolution PC Software 12
Instrument Data Output 12
Performing a Measurement 12
USER INTERFACE 13
Keypad 13
Instrument Firmware 13
Home Screen 14
Applications 14
Single Wavelength 14
Concentration 16
Wavescan 18

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TABLE OF CONTENTS
5061-058-REV-11
Kinetics 19
Standard Curve 21
Multi Wavelength 24
Absorbance Ratio 26
Favourites, Methods, and USB Memory Stick 28
Settings 30
Date and Time 30
Regional 31
Printer 31
Preferences 31
Contrast 32
About the Instrument 32
Additional Options 33
Status Bar Icons 35
USEFUL CALCULATIONS 36
Beer-Lambert Law 36
TROUBLESHOOTING 37
BUILT-IN PRINTER 38
Built-in Printer Accessory Part Numbers 38
Printer Installation Guide
Relling the Printer Paper 39
ORDERING INFORMATION 40

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WARRANTY AND REPAIR
Warranty Policy
IN NO EVENT SHALL BIOCHROM BE LIABLE
FOR INCIDENTAL OR CONSQUENTIAL DAMAGES1
THERE ARE NO IMPLIED WARRANTIES OF
MERCHANTABILITY, FITNESS FOR A PARTICULAR
USE, OR OF ANY OTHER NATURE.1Biochrom
shall not be liable for any claims of any kind whatsoever,
as to the equipment. Warranty is void if the instrument
is modied, disassembled, repair carried out using
unauthorized parts or by a service engineer not
previously approved by Biochrom.
Returns
If any defect arises within or outside the warranty period,
please contact:
US Oce Technical Support
Email: [email protected]
Telephone (Toll Free, US only): 800-272-2775
Telephone (Outside the US): 508-893-8999
Address:
84 October Hill Road
Holliston, Massachusetts 01746, USA
UK Oce Technical Support
Email: [email protected]
Telephone: [phone]
Address:
1020 Cambourne Business Park
Cambourne, Cambridge, UK, CB23 6DW
Goods will not be accepted for return unless the RMA
(Return Materials Authorization) number has been
issued. The unit must be returned with the completed
RMA forms and the Decontamination checklist. The
customer is responsible for shipping charges unless
the failure is within 30 days of receiving the goods.
Please allow a reasonable period of time for completion
of repairs or replacement.
Caution Notice
The Biochrom Lightwave systems are intended for
laboratory use only and can be used in research and
development applications. These systems have been
designed to meet the standards for electromagnetic
compatibility (EMC) and safety intended for laboratory
equipment applications. This product should not be
used in the presence of a ammable atmosphere
such as an anesthetic mixture with air, oxygen, or
nitrous oxide.
1 Where the territory does not allow this exclusion or limitation, this term
will not apply.

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SAFETY INFORMATION
Hazards and Warnings
This section describes potential hazards which may
exist in the operation of these units. A number of
warning labels and symbols are axed to your
instrument. These symbols are used to inform you of
potential dangers which may exist or where caution
is required. Before installing your new unit, please
take time to familiarise yourself with these warnings
and symbols.
This instrument is subject to the following
identied hazards:
This unit uses a Xenon lamp. The lamp energy is
mainly conned within the unit but traverses the
cell holder when a measurement is being taken.
Although the energy present is low and intermit-
tent you are advised not to stare into the beam
or attempt to deect the beam as prolonged
exposure could result in permanent eye damage.
High voltages exist within the power supply unit
and the Xenon lamp housing. Repair and main-
tenance should only be carried out by individuals
trained to work on these instruments.
There are no biohazardous materials within
the unit, however, this unit may be exposed to
biohazardous samples during normal laboratory
use. To protect users against these hazards we
recommend the following decontamination
procedures:
• Wipe the exterior casework with disinfectant
cleaning wipes.
• Remove cuvettes and cuvette holders.
• Wash with disinfectant appropriate
for the biohazard in question.
• Rinse with distilled water.
• Allow to dry thoroughly before reuse.
To further reduce the possibility of biohazards:
• Include an appropriate decontamination
certicate for equipment returned for repair.
• Ensure that the operator of the equipment is
provided with a safe working environment.
• Use, store and dispose of any chemicals in
accordance with manufacturer’s guidelines
and local safety regulations.
• Provide suitable ventilation when working
with volatile solvents or toxic substances.
• Dispose of solvents and chemicals that may
be classed as hazardous waste in accor-
dance with local regulatory practice.
• Determine if personal protective equipment
(PPE) is required for handling laboratory
samples.
All models can be connected to and operat-
ed from a PC. To preserve the integrity of the
measuring equipment it is essential that the
attached PC itself conforms to basic safety
and EMC standards and is set up in accor-
dance with the manufacturers’ instructions.
If in doubt, consult the information that came
with your PC.
The following safety precautions should be
observed when operating a PC:
• To reduce the chance of eye strain, set up
the PC display with the correct viewing
position, free from glare and with appropriate
brightness and contrast settings.
• To reduce the chance of cross contamination
from biological samples, use appropriate
personnel protection measures and
disinfectant wipes on keyboard and mouse.
In the event of contamination, malfunction
or hazard occurring, the operator should
disconnect the unit, by removing the power
cord, and isolate for decontamination
and/or repair.

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INTRODUCTION
Biochrom WPA Spectrophotometers
Spectrophotometers are ubiquitous among modern laboratories. Ultraviolet (UV) and Visible (VIS)
spectrophotometry has become the method of choice in most laboratories concerned with the identication
and quantication of organic and inorganic compounds across a wide range of products and processes.
Applied across research, quality, and manufacturing, with continuing focus on life science and pharmaceutical
environments, they are equally as relevant in agriculture, animal husbandry and shery, geological exploration,
food safety, environmental monitoring, and many manufacturing industries to name a few.
The WPA Lightwave spectrophotometers are quick, accurate, and reliable. They require only small demands
on the time and skills of the operator. This operating manual details the processes for taking basic measurements
using the Lightwave 3 and Lightwave 3+ spectrophotometers.
The Lightwave 3 instrument is UV-VIS split-beam spectrophotometers with a 5 nm spectral bandwidth and
a 40 mm pathlength cell holder. The hardware of the Lightwave 3+ model variants dier only by having a
narrower 3 nm spectral bandwidth.

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INSTALLATION
Unpacking
• The unit weighs less than 4 kg. No special handling is required.
• Please keep the original packaging for transport for service or repair as it has been specically designed to
protect the unit from damage during transit.
• Inspect the instrument and its power supply for any signs of damage caused during transit. If any damage
is discovered, do not use the instrument and report the problem to your supplier.
Positioning
• Ensure your proposed installation site conforms to the environmental conditions for safe operation:
• Indoor use
• 5°C to 40°C
• Maximum relative humidity 90% up to 31°C decreasing linearly to 50% at 40°C.
• Extremes of temperature may require recalibration of the unit for optimal performance.
• The instrument must be placed on a stable, level bench or table capable of supporting its weight allowing
sucient space around the instrument for air to circulate freely.
• The instrument should be positioned so that the power supply cable may be readily removed in the event
of a hazard or malfunction.
• Locate the instrument in an atmosphere free from dust and corrosive fumes. Use the dust cover to further
protect the instrument when not in use.
Installing
• If the instrument has been stored in a cold environment, it should be allowed to come to room temperature
before turning it on to avoid compromising the internal calibration procedure.
• The equipment is operated using an 18 VDC power supply adapter unit. Always use the power supply
adapter and mains cords supplied with the instrument.
• Mains power requirements are as follows:
• 100 to 240 VAC~
• 50 or 60 Hz
• The UK style mains cord plug has a user replaceable
3A fuse. Replace only with the same rating
and type 3A BS1362.
• The unit maximum power rating is 40 VA.
• Connect the instrument to the mains power using
the main power cord and the 18 VDC power
supply adapter unit.
• Switch on the instrument after it has been plugged
in. The instrument will perform a series of self-
diagnostic checks. 18 VDC power supply socket

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INSTRUMENT OVERVIEW
Scope
This user manual covers the following range of WPA UV/Visible spectrophotometers:
Part Number Description
80-3007-42 WPA Lightwave 3
80-3007-43 WPA Lightwave 3 with Printer
80-3007-44 WPA Lightwave 3 with Bluetooth
80-3007-45 WPA Lightwave 3 with Printer and Bluetooth
80-3007-47 WPA Lightwave 3+
80-3007-48 WPA Lightwave 3+ with Printer
80-3007-49 WPA Lightwave 3+ with Bluetooth
80-3007-50 WPA Lightwave 3+ with Printer and Bluetooth
Spectrophotometer Principle and Intended Use
UV/Visible spectrophotometers measure the transmission of light through a sample. Samples absorb light
based on their unique molecular composition. The amount of absorbance is directly proportional to the sample
concentration and the pathlength, which is the distance that the light travels through the sample.
The WPA Lightwave spectrophotometers are intended for laboratory use to measure the concentration of nucleic
acids and proteins in experimental samples.
Hardware
Your spectrophotometer is a simple-to-use UV/Visible instrument with a CMOS array detector (1024 pixels).
It has no moving parts, which is the basis of the rapid scanning operating system. The look and operation
of the Lightwave 3 and 3+ are identical; the only dierence between them is the bandwidth. Throughout the
rest of this manual, the term Lightwave 3 will be used to cover both instruments.

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INSTRUMENT OVERVIEW
Technical Specications
Wavelength Range 190 to 1100 nm
Monochromator Flat grating
Wavelength Calibration Automatic upon switch on
Beam Height 15 mm
Spectral Bandwidth 5 nm or 3 nm for ‘+’ models
Wavelength Accuracy ±2 nm
Wavelength Reproducibility ±1 nm
Light Sources Xenon ash lamp
Detector Twin CMOS array
Photometric Range -0.300 to 2.500 A, 0.3 to 199 %T
Photometric Linearity ±1.3 % or ±0.008 A whichever is greater at 546 nm
Photometric Reproducibility ±0.002 A to 0.5 A at 546 nm
Stray Light <0.5 %T 340 nm
Stability ±0.01 A/h at 340 nm
Noise ±0.005 peak to peak ± 0.002 RMS
Digital Output USB Flash Drive, PC via PVC software, Optional Bluetooth
Data Export USB Flash Drive: .tsv, native PVC format
PC via PVC: .csv, .emf, .xlsx, .xls, .rtf, .tsv, native PVC format
Method Storage 90 with PIN number protection
Sample ID Yes
Languages English, German, French, Spanish, Italian, Japanese, Chinese
Dimensions 260 × 390 × 100 mm
Weight 3.00 kg
Power Input 18 VDC at max 40 VA from a supplied 100 to 240 V~,
50/60 Hz Mains Power Adapter
Display and Keypad
The instrument has a ¼ VGA (320 × 240 pixel) resolution backlit LCD display. The instrument’s built-in
rmware is navigated using the hard-wearing, spill-proof membrane keypad.
Instrument Connections
USB connector for PC connection USB connector for USB memory stick

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INSTRUMENT OVERVIEW
PVC PC Software
The instrument is supplied with the PVC software program (supplied with its own devoted operating manual)
on the accompanying USB ash drive. The instrument can be connected to a PC onto which the PVC
software has been installed, via a USB A to USB B cable or the factory t Bluetooth accessory. This enables
the operator to “print through” the PC directly to the printer that is connected to it. The data may also be
stored as a comma-separated value (.csv), enhanced meta le (.emf), Excel spreadsheet (.xlsx, .xls), rich
text format (.rtf), tab-separated value (.tsv) or in a native PVC format le.
Biochrom Resolution PC Software
When connected to a PC the spectrophotometers can be controlled using the Biochrom Resolution PC
software packages. Operation using Biochrom Resolution PC software is described in the Resolution user
manual or Resolution help le.
Instrument Data Output
A printer accessory is available for the instrument. This may either be supplied preinstalled or as an
optional accessory for end-user installation.
Measurement data can also be exported to a USB ash drive via the USB A socket on the side of the
instrument, as either a tab-separated value (.tsv) or native PVC format le.
Performing a Measurement
The optical height (z value) of the instrument is 15 mm. The light path is directed from RIGHT to LEFT through
the cell chamber.
The cell holder supplied with the instrument accepts standard quartz, glass or plastic cuvettes with pathlengths
up to 40 mm. When using a cuvette with a pathlength less than 40 mm ensure the cell is inserted to the far
right of the cell holder. Cuvettes with a pathlength of less than 10 mm should be secured using an appropriate
packing piece.
12 mm test tubes may be used (for cell cultures for example), however they are not recommended as higher
quality data is produced by using disposable cuvettes. If used, align the indicator line on 12 mm test tubes in
the same direction to ensure reproducible positioning of the tube.
Please consult the “User Interface” section of this user manual for more detail on taking a measurement using
the spectrophotometer. In summary, how to perform a measurement is outline below.
1. Open the desired application on the spectrophotometer.
2. Insert a cuvette containing the reference sample into the cuvette holder.
3. Take a reference measurement using the reference key; the acquired reference baseline is applied to
any subsequent sample measurements until a new reference baseline is taken, or the application is closed.
4. When the reference is complete, remove the reference sample containing cuvette from the cuvette holder,
and replace it with a test sample containing cuvette then take a sample measurement using the conrm key.
5. Repeat step 4 until all the sample data has been collected (see the “Saving and Printing” section for
post measurement options).

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USER INTERFACE
Keypad
The instrument is controlled using the snap-dome switch-keys of the membrane keypad, oering tactile
feedback during operation.
Instrument Firmware
The instrument rmware uses an intuitive menu arrangement that is navigated using the membrane keypad.
Upon powering on the instrument, and after completion of the internal calibration, the home screen is displayed.

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USER INTERFACE
Home Screen
The instrument home screen is the rst screen displayed after initialisation of the instrument, and all applications
and settings can be accessed from here.
Applications
The Applications screen is accessed from the home screen using the ‘1’ key. It contains basic applications
with denable parameters to meet the needs of typical laboratory protocols.
Home screen for the
Lightwave 3 spectrophotometer
Home screen for the Lightwave 3 spectrophotom-
eter displaying the USB memory Stick application,
made available when a USB Flash drive is inserted

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USER INTERFACE
Single Wavelength
The Single Wavelength application is accessed from the Applications screen using the ‘1’ key. It can be used
to performs simple absorbance (A) or % transmission (%T) measurements.
Step 1
Set the wavelength, 190 to 1100 nm, using the left and
right arrow or alphanumeric keys.
Step 2
Press the down arrow, select the mode, “Absorbance”
or “%Transmission”, using the left and right arrow keys.
Step 3
Press the down arrow, set the sample prompt, “Yes”
or “No”, to bring up the sample number option before
each measurement, using the left and right arrow keys.
Step 4
Proceed to the measurement screen by selecting
“OK” using the conrm key.
OR
Return to the Applications screen by selecting
“Cancel” using the escape key.
Step 5
Insert the reference sample then take a reference
measurement using the reference key.
The acquired reference sample baseline will be
applied to all subsequent sample measurements.

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USER INTERFACE
Step 6
Replace the previous sample with a test sample then
take a sample measurement using the conrm key.
Repeat for all samples.
WARNING!
Previous measurements cannot be recalled so should
be printed or output to a USB ash drive before any
subsequent samples are measured.
Results
The results are displayed on the screen. Use the left and right arrow keys to move the cursor and display
the absorbance at ±15 nm either side of the set wavelength.
Return to the Applications screen using the escape key OR display the options menu using the options key
(see the “Additional Options” section).
Concentration
The Concentration application is accessed from the Applications screen using the ‘2’ key. It can be used to
apply a known factor, or one determined using a standard of known concentration to a single wavelength
absorbance (A) measurement.
Step 1
Set the wavelength, 190 to 1100 nm, using the left and
right arrow or alphanumeric keys.
Step 2
Press the down arrow, select the mode, “Factor”,
“Standard” or “Negative Factor”, using the left and right
arrow keys.
For “Factor” or “Negative Factor” mode, press the
down arrow and enter a value of up to four signicant
gures, using the alphanumeric keys.
OR
For “Standard” mode, press the down arrow and enter
a value of up to four signicant gures, for the known
concentration of the standard to be used using the
alphanumeric keys.

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USER INTERFACE
Step 3
Press the down arrow, enter up to 8-digits to dene the
units that the measurements will be reported in.
OR
Open the Units option using the options key:
Select one of the predened units; “µg/ml”, “µg/µl”,
“pmol/µl”, “mg/dl”, “mmol/l”, “µmol/l”, “g/l”, “mg/l”,
“µg/l”, “U/l”, “%”, “ppm”, “ppb”, or “conc”, using the
left and right arrow keys.
Press the down arrow, select the number of decimal
places to display the concentration in, “Auto”, “0”, “1”,
or “2”, using the left and right arrow keys.
Implement the changes and return to the parameters
screen by selecting “OK” using the conrm key.
OR
Reject the changes and return to the parameters
screen by selecting “Cancel” using the escape key.
Step 4
Press the down arrow, set the sample prompt, “Yes”
or “No”, to bring up the sample number option before
each measurement, using the left and right arrow keys.
Step 5
Proceed to the measurement screen by selecting “OK”
using the conrm key.
OR
“Return to the Applications screen by selecting
“Cancel” using the escape key.

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USER INTERFACE
Step 6
Insert the reference sample then take a reference
measurement using the reference key.
The acquired reference sample baseline will be applied
to all subsequent sample measurements.
Step 7
For “Standard” mode, replace the previous sample with
a standard sample then take a standard measurement
using the conrm key.
Conrm the concentration of the standard sample by
selecting “OK” using the conrm key.
OR
Change the concentration of the standard sample by
entering a value of up to four signicant gures, using
the alphanumeric keys.
Implement the change and return to the measurement
screen to run the standard measurement by selecting
“OK” using the conrm key.
OR
Reject the changes, return to the measurement and
cancel the standard measurement run by selecting
“Cancel” using the escape key.

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USER INTERFACE
Step 8
Replace the previous sample with a test sample then
take a sample measurement using the conrm key.
Repeat for all samples.
WARNING!
Previous measurements cannot be recalled so should
be printed or output to a USB ash drive before any
subsequent samples are measured.
Results
The results are displayed on the screen.
Return to the Applications screen using the escape key OR display the options menu using the options key
(see the “Additional Options” section).
Wavescan
The Wavescan application is accessed from the Applications screen using the ‘3’ key. It can be used to perform
absorbance (A) or % transmission (%T) measurements across a range of wavelengths creating an absorbance,
or transmission, spectrum.
Step 1
Set the start wavelength, 200 to 940 nm, using the
left and right arrow or alphanumeric keys.
Step 2
Press the down arrow, set the end wavelength,
210 to 950 nm, using the left and right arrow or
alphanumeric keys.
Step 3
Press the down arrow, select the mode, “Absorbance”
or “%Transmission”, using the left and right arrow keys.
Step 4
Press the down arrow, set the sample prompt, “Yes”
or “No”, to bring up the sample number option before
each measurement, using the left and right arrow keys.

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USER INTERFACE
Step 5
Proceed to the measurement screen by selecting “OK”
using the conrm key.
OR
Return to the Applications screen by selecting “Cancel”
using the escape key.
Step 6
Insert the reference sample then take a reference
measurement using the reference key.
The acquired reference sample baseline will be applied
to all subsequent sample measurements.
Step 7
Replace the previous sample with a test sample then
take a sample measurement using the conrm key.
Repeat for all samples.
WARNING!
Previous measurements cannot be recalled so should
be printed or output to a USB ash drive before any
subsequent samples are measured.
Results
The results are displayed on the screen. Use the left and right arrow keys to move the cursor and display
the absorbance at each wavelength position.
Return to the Applications screen using the escape key OR display the options menu using the options key
(see the “Additional Options” section).

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USER INTERFACE
Step 1
Set the wavelength, 190 to 1100 nm, using the left and
right arrow or alphanumeric keys.
Step 2
Press the down arrow, set the delay time before the
rst measurement, 0 to 600 seconds, using the left
and right arrow or alphanumeric keys.
Step 3
Press the down arrow, set the duration of the
observation, 1 to 60 minutes, using the left and right
arrow or alphanumeric keys.
Step 4
Press the down arrow, set the interval between
individual measurements, 5 to 60 seconds, using the
left and right arrow or alphanumeric keys.
Step 5
Proceed to the second parameters screen by selecting
“Next” using the conrm key.
OR
Return to the Applications screen by selecting “Cancel”
using the escape key.
Step 6
Select the mode to dene the desired nal result,
“Delta A”, “Final A”, or “Slope”, using the left and right
arrow keys.
Kinetics
The Kinetics application is accessed from the Applications screen using the ‘4’ key. It can be used to perform
a series of absorbance (A) measurements over a dened timeframe creating a time-course trace.

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USER INTERFACE
Step 7
Press the down arrow, enter up to 8-digits to dene
the units that the nal result will be reported in.
OR
Open the units options using the options key:
Select one of the predened units; “µg/ml”, “µg/µl”,
“pmol/µl”, “mg/dl”, “mmol/l”, “µmol/l”, “g/l”, “mg/l”,
“µg/l”, “U/l”, “%”, “ppm”, “ppb”, or “conc”, using the
left and right arrow keys.
Press the down arrow, select the number of decimal
places to display the concentration in, “Auto”, “0”, “1”,
or “2”, using the left and right arrow keys.
Implement the changes and return to the parameters
screen by selecting “OK” using the conrm key.
OR
Reject the changes and return to the parameters
screen by selecting “Cancel” using the escape key.
Step 8
Press the down arrow, enter a factor to be used to
generate the nal result of a value of up to four signi-
cant gures, using the alphanumeric keys.
Step 9
Press the down arrow, set the sample prompt, “Yes”
or “No”, to bring up the sample number option before
each measurement, using the left and right arrow keys.
Step 10
Proceed to the measurement screen by selecting “OK”
using the conrm key.
OR
Return to the previous parameters screen by selecting
“Back” using the escape key.