Thermo Scientific Evolution Pro User manual

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Evolution Pro
Spectrophotometer
User Guide
269-350000 Revision A August 2021

WARNING Avoid an explosion or fire hazard. This instrument or accessory is not
designed for use in an explosive atmosphere.
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The contents of this document are subject to change without notice. All technical information in this
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For Research Use Only. This instrument or accessory is not a medical device and is not intended to be
usedfor the prevention, diagnosis, treatment, or cure of disease.

Thermo Scientific Evolution Pro Spectrophotometer User Guide
1
DANGER
Indicates a hazardous situation which, if not avoided, will result in death or
serious injury.
WARNING
Indicates a hazardous situation which, if not avoided, could result in death or
serious injury.
CAUTION
Indicates a hazardous situation which, if not avoided, could result in minor or
moderate injury.
NOTICE
Follow instructions with this label to avoid damaging the system hardware or
losing data.
Note
Contains helpful supplementary information.
Evolution Pro Spectrophotometer
Considerations
Site Preparation and Safety
Before using the system, read the site preparation and safety manual on the documentation
media provided. Always follow the safety precautions in that manual and in this document
when using the system.

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Evolution Pro Spectrophotometer User Guide Thermo Scientific
Operating Precautions
WARNING
• Do not operate this system without following the safety precautions described in this
manual and the documentation that came with your system.
• n'utilisez le système qu'en suivant les consignes de sécurité qui figurent dans ce
manuel et dans la documentation accompagnant le système.
• Betreiben Sie dieses System nur unter Beachtung der Sicherheitshinweise in diesem
Handbuch und in der im Lieferumfang des Systems enthaltenen Dokumentation
• Non utilizzare questo sistema senza seguire le norme di sicurezza descritte in questo
manuale e nella documentazione di accompagnamento del sistema.
• No opere este sistema sin sequir las precauciones de seguridad descritas en este manual
y en la documentación proporcionada con el sistema.
Operating Precautions
The spectrophotometer contains precise optical components. Handle it carefully and follow
these precautions.
• Do not allow moisture to leak into the instrument interior
• Wipe off spilled chemicals immediately
• Do not drop the instrument
• Protect the instrument from mechanical shock
• Protect the instrument from dust
Spectrophotometer Basics
Sample Compartment Lid
Lift the blue tab at the front of the sample
compartment to raise the entire lid.
Slide the door in the lid to the back to provide easy
access to the sample position to insert or remove
a cuvette.
Stainless steel plate to place the samples to
be tested.
Sample
compartment
Sliding
door
Stainless
steel tray

Spectrophotometer Basics
Thermo Scientific Evolution Pro Spectrophotometer User Guide
3
Note
The Z-height (distance from the bottom of the cell to the center of the light beam)
for the spectrophotometer is 8.5 mm. This is true for all cuvette holder accessories. If
purchasing microcuvettes be sure to specify 8.5 mm as the Z-height for the window.
Front Cover
Remove the hold-down screw on the front panel.
Pull upwards to remove it
Align the tabs on the sides with the grooves in the
instrument cover and slide the panel into place to
replace it.
The hold-down screw is generally only required
when tubing or hoses are being brought into the
sample compartment under the flaps at the bottom
of the front cover.
Installing and Removing Accessories
Standard Cell Holder
The standard cell holder included with the
instrument accommodates 10 mm pathlength cells.
Accessories that are not electrically connected are called manual accessories. It is not necessary
to power off the instrument while installing or removing a manual accessory.
Installing a Manual Accessory
1. Open the sample compartment door.
2. Remove any accessory from the location where you plan to install the new accessory (see
the instructions below).
Pin A
Pin A
Finger hole
Reference position cell
holder
Pin B
Sample position
Pin B
Toggle

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Evolution Pro Spectrophotometer User Guide Thermo Scientific
Smart Accessories
3. Position the accessory so the two alignment holes on its baseplate fit over pins A and B in
the sample compartment.
4. Gently push down on the front edge of the baseplate until it snaps in place under the
toggle.
5. Close the sample compartment door.
For operating instructions and installation details, see the user guide for the accessory.
Removing a Manual Accessory
1. Use the handle or finger hole on the accessory baseplate to release the plate from the
toggle.
2. Lift the plate off the alignment pins and remove the accessory from the sample
compartment.
Smart Accessories
Smart Accessories include cell changers and sample holders that feature:
• Auto recognition
• Smart alignment
• Serial number reporting
A connector under the accessory baseplate provides data communication and power. Smart
accessories are hot swappable. It is not necessary to power off the instrument while installing
or removing a Smart Accessory.
Reference connector
Sample connector
These accessories are supported in the sample position:
• Smart 7-cell rotary changer
– Also supports CVC accessory
• Smart 8-cell linear changer
– Also supports 8-cell Peltier accessory
• Smart sipper

Smart Accessories
Thermo Scientific Evolution Pro Spectrophotometer User Guide
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Only the Smart 7-cell rotary changer is supported in the reference position. With this 7 x 7
configuration each cell in the sample position is measured against a unique reference cuvette
in the reference position. This configuration is useful when the different
concentrations/dilutions of matrix, such as buffer solution, are present in a single batch of
samples.
Installing a Smart Accessory
1. Open the sample compartment and remove any cell holder or accessory.
2. Insert the accessory.
a. Grasp the accessory by the handles and lower it into the sample compartment,
aligning the connector under the baseplate with the connector in the sample
compartment floor.
b. Line up the two holes at the back of the accessory with pins A and B in the sample
compartment.
Alignment hole B
Alignment hole A
Pin A
Pin B
D-connector
(female)
c. Press down on the front of the accessory to secure the connection.
The software displays a prompt to initialize the accessory.

Smart Accessories
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Evolution Pro Spectrophotometer User Guide Thermo Scientific
3. Click OK to initialize the accessory.
Initialization reads information about the accessory and, for cell changers, moves it to
position 1.
Refer to the user guide for the accessory for detailed operating instructions.
Removing a Smart Accessory
1. Grasp the handles or the cell holder (if handles are not present) and lift the front of the
accessory to release the baseplate from the toggle.
2. Lift the plate off the alignment pins and remove the accessory from the sample
compartment.
3. The software confirms the accessory has been removed.
Routing Tubing or Cables into the Sample Compartment
The front cover of the sample compartment has two openings to allow passage of tubing or
cables into the sample compartment. These openings are covered by a layer of black foam to
exclude light. Slits cut into the foam allow you to create an opening without allowing excess
light into the sample compartment.
Slits on bottom of panel
CAUTION
• Avoid pinch hazard. Keep hands and objects clear of the accessory during
initialization.
• Évitez tout risque de pincement. Gardez vos mains, ainsi que tout objet, à l’écart de
l’accessoire pendant l’initialisation.
• Quetschungen sind zu vermeiden. Hände und Objekte sind während der
Initialisierung vom Zubehör fern zu halten.
• Evitare il rischio di pizzicamento. Tenere mani e oggetti lontano dall’accessorio
durante l’inizializzazione.
• Evite riesgos de atrapamiento. Mantenga manos y objetos lejos del alcance del
accesorio durante la inicialización.

Smart Accessories
Thermo Scientific Evolution Pro Spectrophotometer User Guide
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Ensure that the water flows freely through the circuit (hoses are not pinched) and that the
tubes do not obstruct the light beam or movement of a cell changer accessory. If the tubing
used has insulation around it, cut the insulation off at a suitable point to allow the section of
tubing that will reside in the opening to be uninsulated.
• For the PCCU1 accessory, allow the portion of the tubing that will be inside the sample
compartment to remain uninsulated. This will not impact performance. See photographs
below.
• For the TPS-1500W or any similar thermostatted recirculator using insulated tubing,
replace the insulation around the tubing inside the sample compartment.
The hold-down screw at the center of the sample compartment front cover should be secured
to hold the cover firmly in place when tubing or cables are present.
Reference Cell Holder
For measurements with a reference sample, install a cuvette containing a suitable reference
solution in the reference position. The reference position can accommodate all manual cell
holders and the Smart Rotary 7-Cell Changer. It cannot accommodate the Linear 8-Cell
Changers or the Smart Sipper.
Connections on the Side of the Instrument
There are three communications cable connections on the right side panel of the instrument.

Using the Spectrophotometer
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Evolution Pro Spectrophotometer User Guide Thermo Scientific
Tip
The power light flashes slowly when the spectrophotometer is starting up. When the
start-up sequence is complete the light will be steady. If the power light starts to flash
rapidly this indicates an error condition. Check INSIGHT software for any error reports.
If INSIGHT it not running or shows no error
1. Power the spectrophotometer OFF.
2. Perform basic checks and prepare to collect fault information:
a. Check that both sample and reference beams are clear of obstacles.
b. Close the sample compartment doors.
c. Start INSIGHT software on the attached computer.
d. Power the spectrophotometer ON.
e. Watch the progress of the startup routine.
3. If the error repeats, note the stage (bottom left corner of the window) where the error
occurred and contact us.
• Trigger port – connect a contact closure trigger device here
– Used with the rapid mix kinetics accessory
– Used with third party remote triggering devices
• Accessory Port – provides support for planned future accessories
• USB-B Port – connect the computer running the instrument control software here
Using the Spectrophotometer
Connecting and Powering the Spectrophotometer
Refer to the Getting Started guide supplied with the instrument (printed copy) and also on
the same media as this user guide.
Making a Measurement
Refer to the INSIGHT software help system, included with your installation of INSIGHT
software. INSIGHT software help includes quick-start guides for each application mode.
These are available with one click from the help system home screen.
The beam direction in the sample compartment is from right to left.
The beam direction in the reference compartment is from front to back.

Using the Spectrophotometer
Thermo Scientific Evolution Pro Spectrophotometer User Guide
9
Note
A question commonly asked of our technical support specialists is “I have a cuvette
with blank solution in the reference beam. Why do I still need to place a cuvette with
blank solution in the sample position and record a blank or baseline? Why can’t I just
measure my sample directly?
The short answer is “That is not how the reference channel works.” A “blank”
measurement is required so that the instrument knows what “100%T” looks like on the
sample detector. The 100%T value is required in order to calculate %T for the sample.
That value is required in order to calculate absorbance.
Most cuvettes have two frosted walls and two polished walls. Try not to touch the polished
walls. Wipe the polished walls with a lint-free cloth to remove dust, droplets of liquid or
fingerprints before making a measurement.
Inserting and Removing Cuvettes
Always insert cuvettes so that the beam passes through the polished walls. Plastic cuvettes
generally have a “V” or other mark at the top of one polished wall. Ideally, you should always
orient this “V” in a consistent direction relative to the beam. Generally, this is towards the
beam.
Using the Reference Beam
For simple measurements it is not generally necessary to put a reference cuvette containing
blank solution in the reference beam. There is no need to “balance the beams” in an
Evolution Pro spectrophotometer. A cuvette in the reference beam is only necessary if:
• You expect the matrix (for example, a buffer solution) to change absorbance during the
course of the measurement. This typically only applies to kinetics (Rate) experiments that
last several minutes or longer.
• Where the method calls for placing a reagent blank in the reference beam as part of the
protocol that will effectively subtract out the absorbance of the reagent blank.
• Where an existing SOP or protocol developed for an older instrument has been validated
and cannot be deviated from. It will do no harm to place a cuvette with blank solution in
the reference beam.

Lamp Compartment Access Panel
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Evolution Pro Spectrophotometer User Guide Thermo Scientific
NOTICE
The warranty does not cover damage to internal optics or electronics caused by
failure to use sample compartment windows or to adequately purge the sample
compartment.
Lamp Compartment Access Panel
A mercury lamp accessory is available for the Evolution Pro either as a factory-installed
option or as a user (or Field Service Engineer) installed accessory. To access the lamp
compartment, turn the locking screw 90 degrees counterclockwise and lift the panel upwards.
Corrosion Protection
The spectrophotometer contains precise optical components that may be damaged by a
corrosive environment.
• Quartz windows on the monochromator cover protect the monochromator from
corrosive gases in the sample compartment to some extent. However, the monochromator
is not sealed and corrosive gases from the sample compartment can still make their way
into the monochromator through other routes.
• There are no windows between the detector compartment and the sample compartment.
If your samples will produce corrosive vapors you should purge the sample compartment with
a continuous flow of draw gas such as dried air or nitrogen.
Purging the Sample Compartment
Purging the sample compartment removes moisture from the sampling area for subambient
temperature-controlled measurements. It also serves to flush potentially harmful vapors from
volatile samples out of the sample compartment.

Corrosion Protection
Thermo Scientific Evolution Pro Spectrophotometer User Guide
11
Note
Ensure the tubing does not obstruct the light beam or prevent the sample
compartment door from closing. If necessary, secure the tubing using cable clips or ties.
Use dry air or dry nitrogen to reduce or eliminate condensation. The purge gas must be free of
moisture, oil and other reactive materials. To remove particulate matter and oil, install a
10-micrometer filter. Connect the flow of purge gas to the 1/4-inch internal diameter tubing
purge fitting on the back of the spectrophotometer.
The Evolution Pro does not feature a dedicated purge port. Route the tubing for the purge
gas into the sample compartment through one of the openings in the baseplate in the well
beneath the reference position. A flow rate of 17 liters per minute is recommended to
completely replace and exclude room air from the sample compartment.
Installing a Liquid Thermostatted Accessory
Liquid thermostatted accessories have ports to attach hoses from an external thermostatted
recirculator. The heated or cooled liquid flows through internal chambers within the metal
block where the cells are held, bringing that block to the same temperature as the liquid. The
cuvettes then come to the same temperature as the block.
❖ To install a thermostatted accessory
1. Install the accessory in the sample compartment.
2. Route the tubing from the recirculator through the two openings at the base of the
sample compartment front cover.
See “Routing Tubing or Cables into the Sample Compartment” on page 7.
3. Use additional tubing and adapters (not included), if necessary, to size the tubing from
the recirculator to match to the ports on the accessory.
For a cell changer, set the changer to position 1 before connecting the tubing to the ports on
the accessory. Allow just enough tubing inside the sampling compartment to permit the cell
changer to move through all possible positions
WARNING
• Do Not use a flammable gas to purge the instrument
• N'utilisez jamais de gaz inflammable pour purger l'instrument
• Verwenden Sie niecontact us
• mals entflammbares Gas zum Spülen des Instruments
• Non utilizzare mai un gas infiammabile per spurgare lo strumento
• Nunca use un gas inflamable para purgar el instrumento

Cleaning the Instrument
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Evolution Pro Spectrophotometer User Guide Thermo Scientific
NOTICE
Do not allow moisture to leak into the instrument interior.
Cleaning the Instrument
Clean the instrument regularly using approved methods (wipe off spilled chemicals
immediately).
1. Use a lint-free cloth dampened with a weak solution of detergent and water to wipe the
surface as necessary.
2. Repeat using a cloth dampened with plain water.
3. Dry the surface with another cloth.
Lamp Replacement
The xenon flash lamps used in Evolution spectrophotometers are extremely long lived. Each is
rated for service equivalent to continuous flashing for a period of three years and is covered
under warranty by us for three years. Under typical operating conditions the lamp will not
require replacement within ten years.
INSIGHT software tracks lamp use and reports remaining lamp life on the Lamp tab in the
System Settings.
Re-alignment of the instrument optics is required following replacement of the lamp. This
means that the process must be completed by a trained and equipped service engineer, either
in the field or at one of our service depots.
Live Readings
In spectrophotometers with traditional, short lived, tungsten or deuterium/tungsten sources
the lamp is always on. It is common for the software running these instruments to display a
value for the absorbance even when the instrument is not actively running an experiment.
The xenon flash lamp is only on when actively making a measurement, so no such display is
present in our software. If you wish to monitor live absorbance readings you can do this using
the Live Display mode in INSIGHT software. Refer to the INSIGHT software user guide or
the Help system within the INSIGHT software for details.