Nano Temper Prometheus Panta User manual

Prometheus Panta
User manual

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Table of contents
1. About this user manual......................................................................................................................... 5
Directions for more detailed information.................................................................................... 5
2. Safety information................................................................................................................................ 6
Symbols and descriptions ............................................................................................................ 6
Use and misuse............................................................................................................................. 7
Safety instructions........................................................................................................................ 7
3. The Prometheus Panta system ............................................................................................................ 9
General.......................................................................................................................................... 9
3.1.1. Intended use ................................................................................................................................. 9
3.1.2. Conformity .................................................................................................................................... 9
3.1.3. Identification................................................................................................................................. 9
Technical information ................................................................................................................ 10
3.2.1. Technical specifications ............................................................................................................. 10
3.2.2. Connections for input and output.............................................................................................. 11
Legal............................................................................................................................................ 12
Limited warranty ........................................................................................................................ 13
4. Prometheus Panta setup.................................................................................................................... 14
Scope of delivery......................................................................................................................... 14
Unpacking................................................................................................................................... 14
Startup ........................................................................................................................................ 14
Cleaning ...................................................................................................................................... 14
Software updates........................................................................................................................ 15
5. Prometheus Panta instrument........................................................................................................... 16
Temperature Range.................................................................................................................... 16
Capillary Format ......................................................................................................................... 16
6. Using Prometheus Panta.................................................................................................................... 17
General Usage............................................................................................................................. 17
Sample Loading.......................................................................................................................... 17
6.2.1. Sample Loading for the Prometheus Panta .............................................................................. 17
7. Patents and intellectual property...................................................................................................... 20

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8. Transport and disposal....................................................................................................................... 21
Repackaging for transport.......................................................................................................... 21
Waste disposal ............................................................................................................................ 21
System disposal.......................................................................................................................... 21

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1. About this user manual
This manual is a guide for using Prometheus Panta instrument. It instructs first-time users on how to use
the instrument and serves as a reference for experienced users.
Before using the Prometheus Panta instrument, please read this instruction manual carefully, and make
sure that the contents are fully understood. This manual should be easily accessible to the operator at all
times during instrument operation. When not using the instrument, keep this manual in a safe place. In
case of loss, please contact NanoTemper Technologies customer support
(https://nanotempertech.com/support) for a replacement copy of this manual.
Directions for more detailed information
There are two sources for further, more detailed information on scientific principles and
recommendations for software usage, as well as data interpretation. One is the NanoPedia knowledge
base that is integrated into the PR.Panta Control software. It contains a variety of articles about the
biochemistry of protein stability, the implemented measurement technologies nanoDSF, backreflection
and DLS and the operational functions of Prometheus Panta. A PDF version of the NanoPedia can be
accessed through the NanoTemper Technologies Explorer Community.
The NanoTemper Technologies Explorer Community is an online community to obtain resources for
NanoTemper products such as application-notes, tech-notes or protocols. It is also a place to exchange
best practices directly with other Prometheus users. You also have the possibility to ask questions and
have them answered by NanoTemper support guides. Follow https://nanotempertech.com/be-an-
explorer/ for access to the Explorer Community.

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2. Safety information
To ensure operation safety, this system must be operated correctly. Carefully read this chapter to fully
understand all necessary safety precautions before operating the system.
Symbols and descriptions
This section describes the safety symbols and descriptions used in this manual, as well as the labels on
the system.
Please take a moment to understand what the signal words WARNING! CAUTION and NOTE mean in this
manual.
WARNING!
AWARNING! indicates a potentially hazardous situation which, if not avoided,
could result in serious injury or even death.
CAUTION
ACAUTION indicates a potentially hazardous situation which, if not avoided, may
result in minor or moderate injury. CAUTION may also be used to alert against
damaging the equipment or the system.
Do not proceed beyond a WARNING! or CAUTION notice until you understand the
hazardous conditions and have taken the appropriate steps.
NOTE
ANOTE provides additional information to help the operator achieve optimal
system and assay performance.
Read manual label. This label indicates that you must read the manual before
using the system. This label is positioned on the back of the instrument.
Warning symbol. This symbol indicates a surface that can heat up and cause burn
injuries. This warning label is positioned on the sample tray.
Warning symbol. This symbol indicates a possible risk for hand injuries by crushing
and sharp edges. This warning label is positioned on the sample tray.

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Use and misuse
Use the Prometheus Panta system only after having read and fully understood this user manual. Use the
system only in perfect condition. If the system shows any signs of damage, stop operation and contact
NanoTemper Technologies GmbH.
Do not modify the system in any way. Do not use it for anything other than its intended purpose.
Safety instructions
CAUTION The Prometheus Panta has to be installed in a way that does not hinder access to the external
power supply and its power plug.
CAUTION Connect the Prometheus Panta instrument power supply in a way that avoids tripping hazards.
CAUTION Do not use extension cords. Damaged cords, plugs or cables need to be replaced immediately.
Failure to comply may result in a risk of personal injury or damage to the instrument.
CAUTION Do not operate the Prometheus Panta instrument with a power supply other than the one
delivered by NanoTemper Technologies.
WARNING! Connect the Prometheus Panta to the AC power supply using the supplied power cable.
The AC power supply, which is connected to the Prometheus Panta, must only be connected to an outlet
that has a ground contact, since the instrument is assembled in line with the specifications for safety
class IEC 61010-1:2010.
WARNING! Danger of electric shock, fire and skin burns. Do not open the system (other than operating
the door via the software). Do not reach into the door opening.
WARNING! Do not operate the Prometheus Panta instrument with substances or under conditions that
pose a risk of explosion, implosion or release of gases. Do not use the instrument with hazardous or
infectious substances.
CAUTION Use only aqueous samples for analysis in the instrument.
CAUTION The Prometheus Panta instrument weighs approx. 35 kg. Two people are required for
transport. Moving the instrument alone entails a risk of personal injury or damage to the instrument.
CAUTION Do not open the instrument manually or anywhere other than the sample loading drawer.
Opening entails a risk of personal injury or damage to the instrument and may only be done by
NanoTemper Technologies staff.
CAUTION Only NanoTemper Technologies staff may service the instrument. Turn off the power switch
and unplug the power cord before servicing the instrument, unless instructed otherwise.
CAUTION The instrument contains a UV-LED. The UV-LED emits invisible ultraviolet radiation (UVB
radiation) when in operation, which may be harmful to eyes and skin, even at brief periods of exposure.

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Do not look directly into the UV-LED during operation. If used as intended, the instrument emits no UV
radiation.
CAUTION Mechanical moving parts within the instrument can pinch or injure your hands or fingers. Do
not touch or open the instrument while parts are moving. Do not reach into the opening when the door is
open.
CAUTION The display pane is made of glass. Broken glass can injure your hands or fingers.
CAUTION The sharp edges of the Prometheus Panta thermal element pose a risk of injury to hands and
fingers.
CAUTION The instrument contains a temperature regulator to control the sample temperature. Some
accessible parts of the instrument can reach temperatures of up to 60 °C. Don’t touch the temperature-
controlled parts of the instrument when the temperature controller is set to high temperatures.
CAUTION Do not use the instrument at ambient temperatures below 15 °C.
CAUTION Use the instrument only at noncondensing conditions (20-70 % humidity, 15-30 °C). At very
high humidity levels, even normal operating temperatures may result in condensation and corrosion.
CAUTION Turn off the instrument when not in use.
CAUTION Do not use 100% ethanol or other types of organic solvents to clean the instrument as they
may remove the instrument paint.
CAUTION The instrument contains a laser module (visible
laser radiation class 3B according to IEC 60825-1: 2014).
Lasers or laser systems emit intense, coherent
electromagnetic radiation that has the potential of causing
irreparable damage to human skin and eyes. Direct eye
contact can cause corneal burns, retinal burns, or both, and
possible blindness. Do not attempt to open the instrument as this poses a risk of personal injury or
damage to the instrument. When the instrument is used as intended it emits laser radiation of LASER
CLASS 1.
CAUTION Remove loose parts (capillaries and capillary lid) before transport as they may damage the
measurement optics
CAUTION The heating element contains nickel. If allergic to nickel wear protective gloves when cleaning
the tray surface to avoid skin irritation and damage.
CAUTION If liquids penetrate the instrument, for example due to spillage, stop operation of the
instrument, disconnect it from the power supply and contact NanoTemper Technologies GmbH.
NOTE Insufficient air supply can cause overheating of the system. Assure enough air supply by not
covering or obstructing the back of the system.
NOTE Do not install the instrument in a location where it may be exposed to direct sunlight.

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3. The Prometheus Panta system
General
3.1.1. Intended use
The Prometheus Panta system provides fast and sensitive determination and quantification of
conformational and colloidal protein stability. For this purpose, it performs nanoDSF (nano-differential
scanning fluorimetry), backreflection, and DLS (dynamic light scattering) measurements in glass
capillaries. The system is intended for research purposes only. It is not to be used for diagnostic
purposes.
3.1.2. Conformity
The following safety and electromagnetic standards were considered:
•IEC 61010-1:2010 Safety requirements for electrical equipment for measurement, control and
laboratory use. Part 1 General Requirements
•IEC 61010-2-010:2014 Safety requirements for electrical equipment for measurement, control and
laboratory use. Part 2-010: Particular requirements for laboratory equipment for the heating of
materials.
•IEC 61326-1:2012 EMC, Electrical equipment for measurement, control and laboratory use –EMC
requirements.
•IEC 60825-1:2014 Safety of laser products.
3.1.3. Identification
The identification labels (Figure 1) are positioned at the rear panel of the instrument. They include
manufacturer information, system model name and serial number (SN), electrical requirements, and the
CE conformity symbol.
Figure 1: Identification labels for Prometheus Panta.

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Technical information
3.2.1. Technical specifications
Electricity
Input external power supply
90–264 VAC ± 10 % 47–63 Hz, 230 VA max
Output external power supply
24 VDC, 11.67 A max
Electrical input
24 VDC, 10 A
Overvoltage category
CAT I
Pollution degree
2
Environmental
Operating temperature
15–30 °C (indoor use only)
Humidity
20-70 %, noncondensing
Operating altitude
max 2000 m
Prometheus Panta dimensions
Width
35 cm (13.8”)
Height
51 cm (20.8”)
Depth
55 cm (20.5”)
Weight
35 kg (77 lbs) net
Power supply dimensions:
Width
21 cm (8.3”)
Height
9 cm (3.5”)
Depth
3 cm (1.1”)
Weight
0.5 kg (1.1 lbs) net max
DLS Laser
Wavelength
405 nm ± 5 nm
Power
100 mW max.
Prometheus Panta Laser classification
The device is LASER PRODUCT CLASS 1
Temperature control
Temperature control range
15 °C –95 °C (at 25 °C)
Optional High Temperature Upgrade
15 °C –110 °C (at 25 °C)
Precision of temperature control
± 0.1 °C

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3.2.2. Connections for input and output
Figure 2: Connections on the Prometheus Panta device.
Type
Function
Ethernet
To connect the system to the Control PC/LAN via Ethernet cable
DC Input
To connect the system to electrical power
On/Off Button
Turning the switch to position “I” switches on the instrument.
All ingoing and outgoing connections can be found on the rear panel of the instrument.
Front
Back
Ethernet connection to host PC
To
Cooling Fans
On/Off Button
Ethernet (LAN)
DC Input
Ethernet (PC)
Capillary Tray Gate
Touch Display

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Legal
1. NanoTemper Technologies shall not be held liable, either directly or indirectly, for any
consequential damage incurred as a result of product use.
2. Prohibitions on the use of NanoTemper Technologies software:
oCopying software for purposes other than backup
oTransfer or licensing of the right to use software to a third party
oDisclosure of confidential information regarding software
oModification of software
oUse of software on multiple workstations, network terminals, or by other methods
3. The content of this manual is subject to change without notice for product improvement.
4. This manual is considered complete and accurate at publication.
5. This manual does not guarantee the validity of any patent rights or other rights.
6. If a NanoTemper Technologies software program has failed, causing an error or improper
operation, this may be caused by a conflict from another program operating on the controlling
PC. In this case, take corrective action by uninstalling the conflicting product(s).
7. NanoTemper and Prometheus are registered trademarks of NanoTemper Technologies GmbH.
8. Unauthorized resale is not permitted.

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Limited warranty
Products sold by NanoTemper Technologies, unless otherwise specified, are warrantied to be free of
defects in materials and workmanship for a period of one year from the date of shipment. If any defects
in the product are identified during this warranty period, NanoTemper Technologies will repair or replace
the defective part(s) or product free of charge.
This warranty does not apply to defects resulting from the following:
1. Improper or inadequate installation.
2. Improper or inadequate operation, maintenance, adjustment or calibration.
3. Unauthorized modification or misuse.
4. Use of unauthorized microwell plates and accessories.
5. Use of consumables, disposables and parts not supplied by an authorized NanoTemper
Technologies distributor.
6. Corrosion due to the use of improper solvents, samples, or due to surrounding gases.
7. Accidents beyond NanoTemper Technologies’ control, including natural disasters.
This warranty does not cover consumables like microwell plates, reagents, labeling kits and the like. It
also does not cover normal wear-and-tear.
The warranty for all parts supplied and repairs provided under this warranty expires on the warranty
expiration date of the original product. For inquiries concerning repair service, contact NanoTemper
Technologies after confirming the model name and serial number of your NanoTemper Technologies
system (see 3.1.3).

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4. Prometheus Panta setup
The Prometheus Panta should be installed by NanoTemper Technologies personnel to ensure safety
measures are taken and to confirm proper functionality of the instrument.
Scope of delivery
Upon receiving the system, please check package contents for completeness. The Prometheus Panta
system package contains the following items:
Item
Description
Prometheus Panta system
-
User manual
This user manual
Power supply and cable
24V power supply, network cable for connection to control PC
Control PC
Laptop for Prometheus Panta system
Unpacking
The Prometheus Panta system should only be unpacked and installed by trained NanoTemper
Technologies personnel to ensure proper functionality of the instrument upon delivery.
Startup
Connect the Prometheus Panta system to power by plugging in the 24V power supply. Connect the
Prometheus Panta system to the control PC using the Ethernet connection at the back of the instrument.
The system starts upon switching the power switch.
Cleaning
The Prometheus Panta system does not need any regular maintenance.
To clean the outside surface of the system, unplug the power supply at the back. Wipe the surface,
including the front display, with a cloth slightly dampened with water or ethanol (70%).
The capillary tray, including its mirrored surface, can be cleaned when the device is switched on and the
capillary tray drawer is in the open position (see 6.1). Clean the mirrored surface of the capillary tray with
a scratch- and dust-free tissue and 99.8 % ethanol prior to experiments.

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In case of broken capillaries on the capillary tray, use a paintbrush or tissue to carefully remove pieces of
glass and to avoid scratching the tray. Other surfaces of the capillary tray can be cleaned in a similar
fashion as the outside of the device.
Software updates
Software updates of the embedded system can only be performed by instructed NanoTemper
Technologies personnel and is part of regular maintenance visits. Software updates of PR.Panta Control
and PR.Panta Analysis software can be performed by the users within the respective software solution.

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5. Prometheus Panta instrument
The Prometheus Panta instrument uses nanoDSF technology to detect changes in the fluorescence of the
amino acids tryptophan and tyrosine over a wide range of temperatures. The instrument is used to
induce thermal unfolding of proteins and to determine thermal unfolding transition temperatures.
Furthermore, thermal protein refolding can be monitored and refolding transition temperatures can be
determined.
Secondly, the Prometheus Panta instrument is equipped with backreflection optics to investigate
aggregation and precipitation in a sample upon heating and to determine aggregation onset
temperatures.
Thirdly, the Prometheus Panta instrument is equipped with DLS (dynamic light scattering) optics to
investigate the particle size and particle size distributions at constant temperature. Furthermore, particle
sizes and size distributions can be measured during thermal unfolding in parallel to nanoDSF and
backreflection to determine onset temperatures of size changes.
Temperature Range
The thermal element inside Prometheus instruments can be precisely adjusted from 15 °C to 95 °C.
Additionally, the instruments can be equipped with a High Temperature Upgrade allowing a maximum
temperature of 110 °C.
Note: If experiments exceed a temperature of 95 °C, or measurement times of 3 hours, capillaries must be
sealed using NanoTemper Technologies Capillary Sealing Paste (PR-P001) and Capillary Sealing Applicators
(PR-P002).
Note: For instruments equipped with a High Temperature Upgrade: When running experiments with a
maximum temperature of 110 °C, the minimum ambient temperature is 20 °C to ensure the instrument
reaches its target temperature.
Note: For measurement temperatures below 20°C, the ambient temperature should be  25°C to ensure the
instrument reaches target temperature.
Capillary Format
The capillary format of the Prometheus Panta is cost-effective, easy to handle and offers maximal
flexibility in the experiment scale, while offering a detection precision that is superior to other
approaches. The number of samples measured in one run can be any number from one to 48 capillaries
or one to 24 capillaries when using capillary chips. Prometheus systems should only be used in
conjunction with high quality glass capillaries or capillary chips provided by NanoTemper Technologies.

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6. Using Prometheus Panta
General Usage
Start the Prometheus Panta instrument using the power switch at the back left of the instrument, turn on
the control computer and start the software (PR.Panta Control). It will take approximately one minute for
the Prometheus instrument to complete hardware initialization. You can use the touch display of the
instrument to set the internal temperature to a desired value, and to open the capillary tray drawer.
These commands can also be performed in the control software.
After the experimental session is finished, turn the Prometheus Panta instrument off using the switch.
Shut down the control computer. No particular sequence needs to be followed.
Note: No equilibration times are required after startup. Prior to temperature ramps, the Prometheus
instruments automatically perform a one minute equilibration routine.
Note: Always remove capillaries and capillary lids prior to transport of the instrument.
Sample Loading
Clean the mirrored surface of the capillary tray with a scratch- and dust-free tissue and 99.8 % ethanol
prior to experiments. Keep the tray surface free from dust, dirt and scratches. 10 µl of sample are
required per capillary for an experiment. Capillaries are dipped into the sample and automatically fill by
capillary forces. Capillary filling is accelerated by holding the capillary horizontally. Make sure that
capillaries are filled completely. Tolerable capillary filling levels for thermal unfolding experiments are
shown in Figure 4 (top right panel). Once capillaries are filled, place them onto the capillary tray and fix
the capillaries by placing the lid onto the tray.
Note: Avoid liquid on the outside of the capillary, since this will alter the fluorescence signal. If there is liquid
on the outside of the capillaries, use a dust- and scratch-free tissue to wipe the capillaries clean.
Note: In case of broken capillaries on the capillary tray, use a paintbrush or tissue to carefully remove pieces
of glass and to avoid scratching the tray.
6.2.1. Sample Loading for the Prometheus Panta
The Prometheus Panta offers maximum flexibility in assay design since it is compatible with single
capillaries as well as capillary chips. The removable capillary frame provides 48 numbered slots for single
capillaries. It is important that the capillaries are positioned in the center of the tray. It is also important
that the meniscus of the sample does not approach the heated capillary tray surface, since this will lead
to rapid evaporation (Figure 3).

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Figure 3: Capillary tray layout and loading for the Prometheus Panta instrument. The removable capillary frame can hold up to
48 capillaries, which are numbered from 1 to 48, starting in the front closest to the user. When filling the capillaries, it is
important that the capillaries are filled completely, and that they are centered on the capillary tray. Once all capillaries are
loaded onto the capillary tray, place the magnetic lid to fix the capillaries.
Alternatively, samples can be loaded into capillary chips. The capillaries on Prometheus nanoDSF Grade
24-Capillary Chips are spaced to enable easy and fast sample loading from 384-well microtiter plates. The
Capillary Chip Filling Station (Cat# NT-AT100) is recommended for manual loading of capillary chips from
microtiter plates since it places the sample at the optimal angle for loading (see Figure 4 B). Correct
positioning of the capillaries on the tray is ensured by the capillary chip format.
Place the chip onto the thermal element with the capillaries facing upward (see Figure 4 A). Use the
number imprinted on the chip for orientation, for example to load your sample number 1. Use the
symbols next to the thermal element for correct chip placement. It is recommended to place the capillary
chip with the imprinted number toward the user, since the capillary closest to the user will be
designated “capillary 1” by the software.
Next, place the lid slowly and evenly on top of the capillary chip (see Figure 4 A). The weight of the lid
holds the capillaries in place. Slight movements of the lid (and the capillary chip) inside of the guiding
structures are normal and will not affect the measurement.
CAUTION: Careless, hasty or diagonal placement of the lid can lead to damage of capillaries or the
thermal element. Keep hands away from the instrument when closing the sample drawer. Especially, be
careful not to get caught between capillary lid and instrument, since this can lead to injury and
instrument damage. If necessary, push the ‘Cancel’ button on the touch display of the instrument to stop
the drawer from closing.

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Figure 4: Manual sample loading of capillary chips. (A) When placed correctly, the plastic chip structure frames the thermal
element and the capillaries rest on top of the thermal element. The metal lid is then placed slowly and evenly on top of the
capillary chip. The red arrow indicates the imprinted number on the capillary chip. (B) The Capillary Chip Filling Station is
recommended for manual loading of capillary chips from microtiter plates.

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7. Patents and intellectual property
Prometheus and the nanoDSF technology are patent protected, especially by the following patents:
US10488326B2, US10545081B2, US10618051B2, US10900879B2, EP2848310B1, EP3150988B1, including
their application and registration in additional countries and jurisdictions.