Harvard Bioscience Biochrom Libra S6+ User manual

5061-071 Rev 1.0
Libra S6+
User Manual
Libra S6+80-5001-20

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Table of Contents
Essential Safety Notes .....................................................................................................................................................4
Unpacking, Positioning, Installation ..............................................................................................................................4
Warranty and Repair ........................................................................................................................................................5
Technical Specifications ……………………………………………………………………………………………………………….6
Operation..……………………………………………………………………………………………………………………………….7
Introduction ……………….…………………………………………………………………………………………………………7
Sample Handling Tips ..……………………………………….……………………………………………………………………7
Keypad and Display ..…………….…………………………………………………………………………………………………8
Software Style …………..……………………………………..……………………………………………………………………9
Applications Folder…………………………………………………………………………………………………………………….10
Applications…………………………………………………………………………………………………………………………11
Single Wavelength…………………………………………………………………………………………...………………11
Concentration…………………………………………………………………………………………………………………13
Wavescan…..………………………………………………………………………………………………………………...16
Simple Kinetics……………………………………………………………………………………………………………….19
Standard Curve……………………………………………………………………………………………………………….22
Multiple Wavelength………………………………………………………………………………………………………….29
Absorbance Ratio…………………………………………………………………………………………………………….31
The Life Science Folder……………………………………………………………………………………………………………….34
Applications (Protein Determination).……………………………………………………………………………………...........35
BCA………………………………………………………………………………………………………………………...…36
Bradford……………………………………………………………………………………………………………………….41
Lowry…………………………………………………………………………………………………………………………..46
Biuret…………………………………………………………………………………………………………………………..51
OD600…………………………………………………………………………………………………………………………55
Methods and Favourties Folder …………………………………………………………………………………………………….57
Saving a Method …………………………………………………………………………………………………………………..57

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Delete a Method …………………………………………………………………………………………………………………..58
Lock Method………………………………………………………………………………………………………………………..58
Unlock Method……………………………………………………………………………………………………………………..59
Favourites…………………………………………………………………………………………………………………………..60
Saving Data onto a USB Memory Stick ………………………………………………………………………………………...61
Utilities Folder ………………………………………………………………………………………………………….……………..63
Date and Time …………………………………………………………………………………………………………………….64
Regional …………………………………………………………………………………………………………………………...64
Export Data ………………………………………………………………………………………………………………………..65
Preference …………………………….…………………………………………………………………………………………..65
Contrast ……………………………………………………………………………………………………………………………66
About ……………………………………………………………………………………………………………………………….66
Print Via Computer ……………………………………………………………………………………………………………………67
Installation ………………………………………………………………………………………………………………………….67
Test Tube and Heated Cell Holder Installation …………………………………………………………………………………….68
Accessories ……………………………………………………………………………………………………………………………69
Cleaning and General Care of the Instrument ……………………………………………………………………………………..69
External cleaning ………………………………………………………………………………………………………………….69
Changing cell holder or removal for cleaning …………………………………………………………………………………..69
Icon Glossary ………………………………………………………………………………………………………………………….70

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Essential Safety Notes
There are a number of warning labels and symbols on your instrument. These are there to inform you where potential
danger exists or particular caution is required. Before commencing installation, please take time to familiarize yourself
with these symbols and their meaning.
Caution (refer to accompanying documents).
Unpacking, Positioning and Installation
Check the contents of the package against the packing list. If any shortages are discovered, inform your supplier
immediately.
Inspect the instrument for any signs of damage caused in transit. If any damage is discovered, inform your supplier
immediately.
Ensure your proposed installation site conforms to the environmental conditions for safe operation:
Indoor use only.
Temperature range 5°C to 35°C. Note that if you use the instrument in a room subjected to extremes of
temperature change during the day, it may be necessary to recalibrate (by switching off and then on again).
Maximum relative humidity of 80% up to 31°C decreasing linearly to 50% at 40°C.
The instrument must be placed on a stable, level bench or table that can take its weight (1.55 kg) to allow air to
circulate freely around the instrument.
This equipment must be connected to the power supply with the power cord supplied. It can be used on 90 – 240 V~,
50-60 Hz supplies.
If the instrument has just been unpacked or has been stored in a cold environment, it should be allowed to come to
thermal equilibrium for 2-3 hours in the laboratory before turning the instrument on. This will prevent calibration failure
as a result of internal condensation.
Switch on the instrument via the keypad ( ) after it has been plugged in. The instrument will perform a series of
self-diagnostic checks.
Please read through this user manual prior to use.
Please contact your original supplier in the first instance if you experience technical or sample handling difficulties.
If this equipment is used in a manner not specified or in environmental conditions not appropriate for safe operation, the
protection provided by the equipment may be impaired and instrument warranty withdrawn.

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Warranty and Repair
Biochrom warrants this instrument for a period of 12 months (1 year) from the date of purchase. Where appropriate,
Biochrom will repair or replace the unit for defects of workmanship or materials. This warranty does not extend to damage
resulting from misuse, neglect, or abuse, normal wear and tear, or accidental damage. This warranty extends only to the
original consumer purchaser.
Products failing within the first 30 days of end user operation are considered dead on arrival (DOA) and where appropriate
a replacement will be given if a repair is not possible. In the instance of a DOA; Biochrom will incur shipping charges of
returning the unit back to the facility.
IN NO EVENT SHALL BIOCHROM BE LIABLE FOR INCIDENTAL OR CONSEQUENTAL DAMAGES. Some states do
not allow the exclusion or limitation of incidental or consequential damages do the above limitation to exclusion may not
apply to you. THERE ARE NO IMPLIED WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR USE,
OR OF ANY OTHER NATURE. Some states do not allow this limitation on an implied warranty, so the above limitation
may not apply to you.
If any defect arises within or outside the warranty period, please contact:
US Office Technical Support
Email [e-mail]
Telephone (Toll Free) 1-800-272-2775
Telephone (Outside the US) 1-508-893-8999
Address 84 October Hill Road
Holliston MA 01746
USA
UK
Office Technical Support
Email [e-mail]
Telephone [phone]
Address 1020 Cambourne Business Park
Cambourne
Cambridge UK
CB23 6DW
Goods will not be accepted for return unless a RMA (Returns Materials Authorization) number has been issued by our
Returns-Repairs Department. The customer is responsible for shipping charges. Please allow a reasonable period of time
for completion of repairs or replacement.

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Technical Specifications
Wavelength range 325nm to 1100nm
Monochromator Flat grating
Wavelength calibration Automatic upon switch on
Spectral bandwidth <7nm
Wavelength accuracy ± 2nm
Wavelength reproducibility ± 1nm
Light sources Pulsed Tungsten halogen
Detector CMOS array
Photometric range - 0.300 to 2.500A, 0.3 to 199%T
Photometric linearity ± 2.0 % or ± 0.010A to 1.000A at 546nm,
whichever is the greater
Photometric reproducibility < 0.002 A at 0A and 500nm
Stray Light < 1%T 340nm
Stability ± 0.005A/h at 0A and 546nm after warm-up
Noise ± 0.002A near 0A and ± 0.020A near 2A at
600nm
Digital output USB memory stick data storage
USB PC connection with PVC software
Data Export PC via USB cable
USB memory stick
Method Storage 90 with PIN number protection
Graphical Display Yes, zoom and track function
Sample ID Yes
Languages English, German, French, Spanish, Italian,
Japanese, Chinese
Dimensions 120 x 230 x 320 mm
Weight 1.55 kg
Power input 18 Vdc at max 18VA from a supplied 100 to
240V~, 50/60 Hz Mains Power Adapter

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Operation
Introduction
Your spectrophotometer is a simple-to-use visible instrument. It has no moving parts, which is the basis of the rapid
scanning operating system. It has been designed to meet the routine spectroscopy needs of customers requiring a
compact instrument that is easy to use. The product is reliable and requires low maintenance.
The user interface is built around folders which are displayed on the home page when the instrument is turned on. After it
is turned on and calibrated, the default home page offers the following choices:
Applications General spectroscopic methods
Favourites A folder to store your more frequently used configured methods
Methods Contains nine folders that can store less frequently used configured methods (nine
methods per folder)
Utilities Instrument set up (date, time, language, printer options, preferences and display settings)
Life Science Standard Life Science methods such as nucleic acid assays, protein assays and cell
counting
The instrument is supplied with a program, PVC (Print via Computer) on the accompanying CD. When used with a USB
cable to connect to a PC onto which the software has been installed, it enables the user to “print through” the PC directly
to the printer that is connected to it. The data may also be stored as an Excel spreadsheet, as an EMF graphics file, a
comma delimited (csv) data file, a tab delimited (txt) data file or in native PVC format for later access.
The instrument has a USB connector located in the front of the unit which allows data to be stored on a USB memory
stick. This data can be stored in a PVC format which can be opened using PVC software or csv data file that can be
imported into Excel.
Sample Handling Tips
The light beam is directed from RIGHT to LEFT through the cell chamber; therefore please ensure the cell is inserted
with the correct alignment.
The cell holder supplied with the instrument accepts standard 10 mm path length quartz, glass or plastic cells.
The optical height is 15 mm

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Keypad and Display
The back-lit liquid crystal display is very easy to navigate around using the alphanumeric entry and navigation arrow keys
on the hard wearing, spill proof membrane keypad.
Key Action
On/Standby
Key Turns the instrument on or back to standby.
Arrow Keys Use the four arrow keys to navigate around the display and select the required
setting for the active (highlighted) option.
View Options Used to select items in a menu or to enter parameters and text descriptions.
Repeated key presses can be used to cycle through lower case, upper case,
and numbers. Wait 1 second before entering the next character. Use the “C”
clear key to backspace and the 1 key to enter a space
Alphanumeric
Keys
Use to enter parameters and to write text descriptions where appropriate or
required. Use repeated key presses to cycle through lower case, upper case,
and numbers. Wait 1 second before entering the next character. Use the “Esc”
key to backspace and to enter a space
Escape/
Cancel Escape from a selection and return to the previous folder. Stop making measurements.
OK Confirm parameters/selection.
Set
Reference
Set reference to 0.000A or 100%T on a reference solution at the current
wavelength in the measurement mode selected. When in scan mode, do a
reference scan.
Take a
Measurement Take a measurement.
Alphanumeric Keys
On/Standby
Key
Arrow Keys
View Options
Escape/Cancel
Set Reference
Take a Measurement
Red light will appear
when USB is inserted
OK Key

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Software Style
The user interface is built around having folders of files which are displayed on the home page when the instrument is
switched on. Different folders are numbered and opened by using the associated number key on the keypad.
Folder Keypad Number Description
1Single wavelength, Concentration, Wavelength scan, Kinetics, Standard Curve, Multiple wavelengths and
Ratio.
2Saved User selected and configured methods
(maximum of 9 methods).
3Sub folder selection for User selected and configured methods (9 sub folders each, containing a maximum of
9 methods).
4Instrument set up (date, time, language, data output,
preferences and display settings).
5 Proteins and Cell counting.

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TheApplications Folder
Options
Within each application the user has the possibility to select various options that define the way results are treated. If not
using a stored method, it is advisable to check that these Options have been appropriately set for your experiment when
coming to the instrument. Note that setting the “History” parameter to on (see Preferences later) will cause the instrument
Function Keypad Number Description
1Absorbance or %T (transmission) at a single user defined
wavelength.
2Concentration measurement at a single wavelength based on
a simple Factor entered or calculated from a single standard.
3Wavelength scan between two user-defined wavelengths. Range 330-950 nm, with user configurable peak finding
function.
4Absorbance versus time measurements either rate or end
value based.
5Generation of calibration curve by measuring standards at a
single wavelength.
6Absorbance or %T (transmission) at up to 5 user defined
wavelengths.
7Ratio of absorbance values at two user specified
wavelengths.

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Step 1
Set wavelength by using keypad numbers or left and
right arrows.
Press the down arrow key.
Step 2
Select the mode, Absorbance or %T, using the left and
right arrows.
Press the down arrow key.
Step 3
Select whether or not to adjust the name of your
sample before each new run, using left and arrow keys
to select Yes or No.
Step 4
To enter the results screen with the selected
parameters press OK or Cancel the selections
and return to the Applications Folder by pressing
Cancel .
Step 5
Insert the reference. Press the reference key .
This will be used for all subsequent samples until
changed.
Step 6
Insert sample and press .
Repeat step 6 for all samples.
to store its last settings. If the “History” parameter is turned off, all parameters and options will return to their default
settings when you leave that application. (Unless it has been saved as a method).
Applications
Single wavelength
This makes simple absorbance (A) and % transmission (%T) measurements on samples, measuring the amount of light
that has passed through a sample relative to a reference (this can be air).
The procedure is as follows:

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Results Screen
The result at the selected wavelength is displayed on
screen.
Use the left and right arrows to move the cursor and
display the value at the cursor position (+/- 15nm from
set wavelength).
Press Cancel to return to the Applications
Folder.
Press to display available options which are
described below.
Options (select using key pad numbers)
1. Return to parameters screen (Step 1 above).
2. Print result via selected method.
3. Toggle between Absorbance and %T mode.
4. Print graph – greyed out if no data are available.
7. Sample number – add a prefix to the sample
number and reset the incrementing number to the
desired value.
8. Save method – use the left and right arrows to
select a folder to store in (Favourites/Methods 1-9),
press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing or wait.

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Step 1
Set wavelength by using keypad numbers or left and
right arrows.
Press the down arrow key.
Step 2
Select the mode, Factor (user entered), Standard
(factor is calculated from a calibration sample), or
Negative Factor (user entered, without a sign) using
the left and right arrows.
Press the down arrow key.
Step 3 (if Factor is selected)
Enter the Factor using the keypad numbers. Range
0.001 to 9999. Use the C button to delete the last digit
entered.
Press the down arrow key.
Step 3 (if Standard is selected)
Enter the concentration using keypad numbers. Range
0.01-9999. Use the C button to delete the last digit
entered.
Press the down arrow key.
Step 4 (if Negative Factor is selected)
Enter a negative factor, without a sign, using the
keypad numbers. Range 0.001 to 9999. Use the C
button to delete the last digit entered.
Press the down arrow key.
Step 5
Units: The user can enter a text string up to 8
characters long. To access a list of pre-defined units
press the Options key and then use the
left/right arrows (µg/ml, µg/µl, pmol/µl, mg/dl, mmol/l,
µmol/l, g/l, mg/l, µg/l, U/l, %, ppm, ppb, conc or none).
These units can also be edited once OK is pressed.
This screen also allows the number of displayed
decimal points (DP) to be selected, from 0 to 2. Note
that the result will always be fixed to 5 significant
figures regardless of how many decimal points are
selected (so 98768.2 will display as 98768 even with 1
decimal point selected). Press OK to store the
chosen parameters or Cancel
.
Concentration
This makes simple concentration measurements on samples, by measuring the amount of light that has passed through a
sample relative to a reference (this can be air). Concentration is obtained by multiplying the measured absorbance at a
specific wavelength by a factor. The factor may be known in advance, or may be calculated by the instrument by
measuring a standard of known concentration.
The procedure is as follows:

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Step 6
Select whether or not to adjust the name of your
sample before each new run, using left and arrow keys
to select Yes or No.
Step 7
To enter the results screen with the selected
parameters press OK or Cancel the selections
and return to the Applications Folder by pressing
Cancel .
Step 8 (if using a Factor or using a Negative Factor
Mode)
Insert the reference. Press the reference key .
This will be used for all subsequent samples until
changed.
Step 9
Insert sample and press .
Step 10 (if using Standard mode)
Insert the reference. Press the reference key
.This will be used for all subsequent samples until
changed.
Press to display the Run Standard screen.
Run the standard by pressing or press cancel
to return to the measure screen.
Step 11
Insert the sample and press .
The concentration of the sample is displayed. Results
shown as ---- indicate the concentration is out of range.
Repeat step 11 for all samples.

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Press to return to the Applications Folder.
Press to display available Options which are
described below.
Note: If below zero or negative the concentration
value calculated will be shown as ---. A measured
negative absorbance (reference measurement
higher absorbance than measured sample
absorbance) would require a negative factor to
create a positive concentration.
Options (select using key pad numbers)
1. Return to parameters screen (step 1 above).
2. Print result via selected method.
3. Toggles on/off, displaying a graph of wavescan +/-
20 nm from selected wavelength.
4. Return to Run Standard screen.
7. Sample number – add a prefix to the sample
number and reset the incrementing number to the
desired value.
8. Save method – use the left and right arrows to
select a folder to store in (Favourites/Methods 1-9),
press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing or wait.

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Step 1
Set start wavelength by using keypad numbers or left
and right arrows.
Press the down arrow key.
Step 2
Set end wavelength by using keypad numbers or left
and right arrows.
Press the down arrow key.
Step 3
Select the mode, Absorbance or %T, using the left and
right arrows.
Step 4
Select whether or not to adjust the name of your
sample before each new run, using left and arrow keys
to select Yes or No.
Step 5
To enter the measurements screen with the selected
parameters press OK or cancel the selections
and return to the Applications Folder by pressing
Cancel .
Step 6
Insert the reference Press the reference key .
This will be used for all subsequent samples until
changed.
Step 7
Insert sample and press .
Repeat step 6 for all samples.
Wavescan
An absorption spectrum can be obtained from your instrument, enabling simple identification of peak height and position.
The procedure is as follows:

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Results Screen
A graph of the wavescan is displayed, along with a
table of Absorbance/%T at each peak. Use the left and
right arrows to move the cursor along the graph. When
it reaches a peak the peak height and width of the peak
is displayed at the top of the screen.
To zoom in on the wavelength scale, use the up arrow.
This auto-scales on the Absorbance/%T scale
(dependent on the Graph Scale option) and this is
retained for subsequent measurements.
To zoom out again, use the down arrow. Press
to return to the Applications Folder.
Press to display available Options.
Options (select using key pad numbers)
1. Return to parameters screen (step 1 above).
2. Print result via selected method.
3. Toggle between Absorbance and %T mode.
4. Displays Peak Detection Parameter Screen. See
description below.
5. Manually adds a peak position to the peak table in
the results screen at the position set by the cursor.
If the cursor is returned to this position the legend
“User Defined Peak” is displayed at the top of the
scan and this option changes to Delete Peak
6. Displays Graph Scale Parameter Screen. See
description below.
7. Sample number – add a prefix to the sample
number and reset the incrementing number to the
desired value.
8. Save method – use the left and right arrows to
select a folder to store in (Favourites/Methods 1-9),
press the down arrow and enter name.
9. Auto-print – toggles auto-print on/off.
Exit options by pressing or wait.
Peak Detection (Shortcut Button 4):
AutoDetect Peaks: Turns on and off the automatic
peak detection. The following options determine how
peaks are detected.
Minimum peak height: Minimum height the peak has
to be above the higher of the two adjacent minima for
the peak to be detected.
Minimum peak width: Minimum width of the peak as
determined by the difference in wavelength between
the higher of the two adjacent minima and the opposing
intersection of that higher minimum level and the peak
profile.

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Peak Detect on Zoom:
Determines whether peaks are
re-assessed and tabulated when the user zooms into a
region of the wavescan. If off, it leaves the peak
detection as determined on the un-zoomed display.
Sort peaks by…: Determines the sequence that
peaks are reported by. Can be wavelength, peak
height or peak width.
Draw Peaks: Switches display of peak cursors on and
off. These show vertical dashed lines displaying the
measured peak height and horizontal dashed lines
showing the peak width
Pressing Cancel ignores the selection,
pressing accepts them.
Add Peak (Shortcut Button 5):
Adds a used defined peak at the current cursor
position. The entry is then displayed in inverse
colouring to discriminate between user defined peaks
and auto-detect peaks. When the cursor is positioned
over the user defined peak a legend “User Defined
Peak” appears at the top of the graph. The option then
changes to Delete Peak to enable the user to remove
the peak.
Note: Storing a method at this stage will save these
user defined wavelengths, each time method is run
Absorbance value at these wavelengths is reported.
Graph Scale (Shortcut Button 6):
This enables the user to set up a defined graph by
defining the limits in either or both of the x and y axes.
Zoom mode:
This sets up the operation of the Zoom keys (up and
down arrows). “x & y axes” expands the display around
the cursor measurement point, whilst the other options
select the absorbance or wavelength axes
respectively. With x or y axis limits set to on, zooming
out will only be permitted to the set limits.
x/y axis limits:
Setting “x (or y) axis limits” to “On” activates the start
and finish points of the desired graph to user defined
specific wavelengths and/or absorbance values.
Pressing Cancel ignores the selection;
pressing accepts them and displays the
required graph.

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Kinetics Parameter 1 Screen
Step 1
Set Wavelength. Enter all numerical values using the
keypad numbers or the left and right arrows.
Press the down arrow key.
Step 2
Enter the delay time in seconds before
measurements are taken. This can be a maximum of
600 seconds (10 minutes).
Press the down arrow key.
Step 3
Enter the duration in minutes over which
measurements are taken. This can be a maximum of
60 minutes.
Press the down arrow key.
Step 4
Enter the interval time in seconds between
measurements using the left and right arrows.
Options are: 5, 10, 20, 30 or 60 seconds.
Step 5
Press Next to go to the next parameters
screen or press Cancel to return to the
Applications Folder.
Kinetics Parameters 2 Screen
Step 6
Select the measurement mode using the left and
right arrows.
Delta A: change in absorbance over the
measurement duration (or selected period).
Final A: absorbance at the end of the measurement
duration (or selected time).
Slope: rate of change of absorbance over the
measurement duration or selected period.
Press the down arrow key.
Simple Kinetics
Kinetics studies, where the change in absorbance needs to be followed as a function of time at a fixed wavelength, can be
readily performed.
Reagent test kits are routinely used for the enzymatic determination of compounds in food, beverage and clinical
laboratories by measuring NAD / NADH conversion at 340 nm. The change in absorbance over a specified time period
can be used to provide useful information when an appropriate factor, defined in the reagent kit protocol, is applied.
Reaction rate and enzyme activity can be calculated if the factor used takes account of the absorbance difference per unit
time, as opposed to the absorbance difference per se.
For this reason, the change in absorbance per minute (ΔA/min), concentration (ΔA/min x factor) and correlation
coefficient (calculated from a best fit of the data points) are displayed. They may not be relevant for simple kinetics
experiments.
The procedure to define a new method is as follows:

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Step 7
Units: The user can enter a text string up to 8
characters long. To access a list of pre-defined units
press the Options key and then use the
left/right arrows (µg/ml, µg/µl, pmol/µl, mg/dl, mmol/l,
µmol/l, g/l, mg/l, µg/l, U/l, %, ppm, ppb, conc or none).
These units can also be edited once OK is
pressed.
This screen also allows the number of displayed
decimal points (DP) to be selected, from 0 to 2 Note
that the result will always be fixed to 5 significant
figures regardless of how many decimal points are
selected (so 98768.2 will display as 98768 even with 1
decimal point selected). Press OK to store the
chosen parameters or Cancel .
Press the down arrow key.
Step 8
Set the Factor by which the result is multiplied to give
the amount in the chosen range using the left and right
arrows. Range of 0.01 to 9999.
Press the down arrow key.
Step 9
Select whether or not to adjust the name of your
sample before each new run, using left and arrow
keys to select Yes or No.
Press Next to enter the Results screen or
press Cancel to return to the Parameters 1
screen.
Results Screen
Insert the reference Press the reference key .
Insert the sample and press to start the run.
Time (min) is displayed at the bottom of the screen,
and absorbance data are plotted on the graph as
testing proceeds. The table below the graph gives:
absorbance values at T0(start of calculation), Tn(finish
of calculation, change in absorbance, slope,
regression parameter (R2) of the calculated slope and
the result calculated from the selected parameter (dA,
final A or slope).
Use the left and right arrows to move the cursor and
display the time and absorbance value at measured
data points.