SOPHION BIOSCIENCE QPatch Compact Manual de usuario

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QPatch Compact User Manual

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Content

1. Introduction to QPatch Compact ...............................................................................4

1.1 Scope and purpose .................................................................................................................4

1.2 Notes and cautions .................................................................................................................4

1.3 System description .................................................................................................................4

1.4 QPlate 8/8X ...........................................................................................................................5

1.4.1 Shelf life of partially used QPlate 8 ..............................................................................................................5

1.5 System architecture................................................................................................................5

1.6 Pipetting light guidance ...........................................................................................................6

1.6.1 Instrument initialization .............................................................................................................................6

1.6.2 QPlate insertion/removal............................................................................................................................6

1.6.3 Active sites on QPlate ................................................................................................................................6

1.6.4 Intracellular (IC) and extracellular (EC) solution addition................................................................................6

1.6.5 Cell addition .............................................................................................................................................6

1.6.6 Compound addition ...................................................................................................................................6

1.6.7 Compound addition for ligand-gated experiments ..........................................................................................7

1.6.8 Cells and compound additions scheme .........................................................................................................7

1.7 Temperature control ...............................................................................................................7

1.8 Sophion Analyzer software.......................................................................................................8

2. Startup and shutdown procedures ............................................................................9

2.1 Startup .................................................................................................................................9

2.1.1 Power up & initialization.............................................................................................................................9

2.1.2 Logon......................................................................................................................................................9

2.1.3 Temperature control ................................................................................................................................10

2.2 Shutdown procedure .............................................................................................................10

3. Conducting an experiment ......................................................................................11

3.1 Terminating a running protocol ...............................................................................................12

4. Operating the system ..............................................................................................13

4.1 Cells ...................................................................................................................................13

4.2 Saline solution requirements ..................................................................................................13

4.3 QPlate barcodes....................................................................................................................13

4.4 Touch screen monitor user interface ........................................................................................13

4.5 Screening station software (SSS) ............................................................................................13

4.5.1 Sophion logo – easy access to manuals and guides in SSS ............................................................................13

4.5.2 Login screen ...........................................................................................................................................14

4.5.3 Left-side menu bar ..................................................................................................................................15

4.1 Operation ............................................................................................................................16

4.1.1 Preflight Check screen..............................................................................................................................16

4.1.2 Maintenance menu ..................................................................................................................................17

4.1.3 Settings menu ........................................................................................................................................18

4.1.4 Setting up the compound map ..................................................................................................................20

4.1.5 Priming the QPlate...................................................................................................................................22

4.1.6 Live view panel .......................................................................................................................................25

4.1.7 Choosing the voltage protocol and run parameters.......................................................................................25

4.1.8 Job finished ............................................................................................................................................28

4.1.9 Results screen ........................................................................................................................................29

4.1.10 Log out screen ......................................................................................................................................29

5. Instrument cleaning and maintenance ....................................................................31

5.1 Daily cleaning.......................................................................................................................31

5.2 Occasional maintenance.........................................................................................................31

5.2.1 Thermoelectric circulator ..........................................................................................................................31

5.3 How to drain the thermoelectric circulator for water...................................................................31

5.4 How to drain the QPatch Compact for water before shipping .......................................................33

5.5 Decontamination before shipping to Sophion Bioscience .............................................................34

6. Troubleshooting ......................................................................................................35

6.1 Issues during instrument startup ............................................................................................35

Problem: QPatch Compact does not power up and the touch screen monitor remains black? ......................................35

Problem: The login screen does not appear after the instrument is turned on? .........................................................35

Problem: I cannot log into the instrument? .........................................................................................................35

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6.2 Issues during instrument use..................................................................................................36

Problem: Liquid accumulates inside the manifold – removal of manifold ..................................................................36

6.3 Issues when running an experiment on the instrument. .............................................................37

Problem: The instrument rejects the QPlate after attempting to read the barcode.....................................................37

Problem: The priming of the QPlate fails .............................................................................................................37

Problem: The priming of some QPlate sites fails...................................................................................................37

Problem: The positioning of cells fails too often....................................................................................................38

Problem: The success rate is consistently lower than expected ..............................................................................38

6.4 Maintenance menu................................................................................................................38

6.4.1 Temperature control menu .......................................................................................................................38

6.4.2 Manifold menu ........................................................................................................................................39

6.4.3 Move to home position .............................................................................................................................39

6.4.4 Move to transport position ........................................................................................................................39

6.4.5 Recover from a jammed/stuck manifold......................................................................................................40

6.5 Conducting a pressure test.....................................................................................................43

6.6 Conducting an artificial reference QPlate (ARQ) experiment ........................................................44

6.7 Priming a QPlate 8/8X ...........................................................................................................45

7. System requirements & specifications.....................................................................47

7.1 Instrument...........................................................................................................................47

7.2 Handling the instrument and precautions .................................................................................47

7.3 Environmental conditions .......................................................................................................47

7.4 Database .............................................................................................................................48

7.5 Sophion Analyzer software .....................................................................................................48

7.6 QPlate 8 ..............................................................................................................................48

7.7 PolyAmp 4 – Patch Clamp Amplifier .........................................................................................48

8. Legal statement ......................................................................................................50

8.1 Copyright.............................................................................................................................50

8.2 Trademarks..........................................................................................................................50

8.3 Compliance ..........................................................................................................................50

8.4 Disclaimer............................................................................................................................50

9. Terms and abbreviations (appendix I) ....................................................................51

10. Declaration of conformity (appendix II)..................................................................53

Sophion Bioscience A/S

Baltorpvej 154,

2750 Ballerup

Denmark

🕽+45 44 60 88 00

Sophion Bioscience, Inc.

213 Burlington Road, Suite 105,

Bedford, MA 10730

United States

🕽+1 877 898 9010

Sophion Bioscience K. K.

Incubation on Campus Honjo Waseda #V203,

1011 Nishitomoda, Honjo-shi,

Saitama 367-0035, Japan

🕽+81 495 71 8811

Sophion Bioscience (Shanghai) Co., Ltd.

Room 550, Floor 5, Building 21

No. 1-28, Lane 588 Tianxiong Road, Pudong District

Shanghai 201318, China

🕽+86 21 52037529

Sophion.com

[email protected]

1. Introduction to QPatch Compact

1.1 Scope and purpose

QPatch Compact (QPC) performs up to 8 independent, high-fidelity voltage clamp or current clamp

experiments at the same time. The system contains a database for data storage and software for

setting up, executing, and analyzing patch clamp experiments.

As an integral part of the QPatch Compact system, this manual must be easily accessible to users.

This user manual is also accessible from the QPatch Compact screening station software (SSS).

Suggestions for improvements are always welcome; please contact us for your feedback at

[email protected].

1.2 Notes and cautions

Throughout the manual, several notes and cautions are mentioned. These are marked in grey, and

orange boxes, respectively. The purpose of notes, and cautions is shown in the following.

Note! A note contains information that should be paid special attention to. Notes

are typically information which is important for correct assay setup. Failure to

conform to notes may negatively affect performance and data quality.

Caution! A caution contains information that should be followed. Cautions are

typically information that is important for correct system setup and use. Failure to

conform to cautions may lead to permanent loss of data, cells, reagents, and

consumables.

1.3 System description

Caution! We recommend that staff familiarize themselves with risks of electrical

measurements, hazardous and toxic liquids, and related biohazards. Users should

read this manual and especially note the “Safety Guide” which is enclosed with the

instrument in the transport case.

QPC is used for measuring ion channel currents or voltages in living cells employing the planar patch

clamp technique. The experiments are performed on silicon chips with either a single or ten-patch

hole. Eight silicon chips are integrated into a QPlate 8/8X.

The QPlate is a ready-to-use consumable (Figure 1). When cells are applied to the QPlate, they will

be moved to the patch hole by a microfluidics system and captured at the patch hole by suction. The

cells to be patched are randomly selected. Therefore, the cells in the sample applied to the patch

clamp chip must be of high quality and uniformly single cell suspended.

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Figure 1 Schematic view of one recording site in a QPlate 8

1.4 QPlate 8/8X

Caution! QPlates must be stored at 4-8°C in the protective bag under vacuum in

which they were shipped. QPlates should not be placed back at 4-8°C after the bag

is opened. This is to avoid condensation on the QPlates, which can disturb the

electrical circuit.

Each measurement site contains two microfluidic flow channels, one for the extracellular and one for

the intracellular solution (Figure 1) and the liquids are driven by fluid pressure. It is important that

both the positive and negative pressure is within the recommended range – see chapter 7.

QPlate 8 are single-hole QPlates and QPlate 8X are the corresponding multi-hole QPlates.

The QPlate 8/8X is a single-use consumable, hence the QPlate recording sites are only once and

cannot be reused.

The extracellular solution is collected in the waste reservoir inside the QPlate 8 and has a volume of

250 µL. It is important to comply with the volume guidance from the system for it to keep track of

the liquid level in the waste reservoir. It is the only container for biological and potentially hazardous

waste and must be disposed of accordingly. Please see our “Safety Guide” which is enclosed with the

instrument in the transport case for more information.

A pair of measurement electrodes are placed at each of the 8 sites and connected to individual

amplifiers.

1.4.1 Shelf life of partially used QPlate 8

Once you start using a QPlate 8, it can be used for one week. When not actually in use, partially used

QPlates 8 should be kept in a desiccator.

1.5 System architecture

QPatch Compact consists of:

•The instrument which handles the QPlate, cells,

compounds, and performs the actual experiments. This

includes the electrothermic circulator which ensures

temperature control.

•The internal Oracle database, used for storing

experimental parameters and measured data.

Figure 2 QPatch Compact

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•The Sophion Analyzer software, used for planning experiments, executing, and analyzing

the data.

The QPC instrument is commanded via the touch screen using the Screening Station Software

(SSS).

The Oracle database stores all recorded data and other information in the system. The database is

the software device used for storage of assay definitions, experiment data and user settings. The

database ensures that the measured data cannot be deleted accidentally. This also applies to all

information on experiment settings and execution. Users must login, and the system records which

users performed which actions in the system. QPatch Compact is delivered with a pre-configured and

pre-installed database which is ready to operate with the instrument.

1.6 Pipetting light guidance

The pipetting light and audio guidance is an assistive feature to the cell and liquid addition steps. It

was incorporated to help guide the user during liquid addition for more accurate timing and a better

user experience.

The light guidance features several functions depending on the particular step in the patch clamp

setup process.

1.6.1 Instrument initialization

During instrument initialization the light guidance will always show an downward-upward flash

sequence. This indicates that the light guidance works as expected.

1.6.2 QPlate insertion/removal

The pipetting light guidance indicates when to insert and remove the QPlates and in which direction

to move the QPlate.

1.6.3 Active sites on QPlate

The active sites which are indicated on the touch screen as the user selects the sites on which to

record are also highlighted with white light next to the manifold.

1.6.4 Intracellular (IC) and extracellular (EC) solution addition

During IC and EC addition, the light guidance shows which sites to pipette into indicated by blue

light.

1.6.5 Cell addition

During cell addition, the light guidance will indicate the next site in which to pipette by a blue light.

The user must wait without inserting the pipette tip into the manifold until the blue light turns orange

while preparing to insert the pipette tip and dispense the liquid. As soon as the blue light turns orange

the user inserts the pipette tip and dispenses the liquid in one clean motion. The next site in which

to pipette will then turn blue and the process repeats with site.

1.6.6 Compound addition

During compound addition, the light guidance will point to the next site in which to pipette indicated

by a blue light like the cell addition step. The user must wait without inserting the tip until the blue

light turns orange. Once it has turned orange the user must insert the tip into the manifold as far as

it will go making sure the tip collar rests on the manifold shoulder and dispense the liquid in one

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clean motion and then remove the tip without aspirating. Please consult our video tutorials “How to

prepare cells for QPatch Compact” and “How to start your experiment on QPatch Compact” available

on our self-learning QPC support site.

1.6.7 Compound addition for ligand-gated experiments

During compound addition, the light guidance will point to the next site in which to pipette indicated

by a blue light similar to cell addition. The user must wait without inserting the tip until the blue light

turns orange.

The washout of the compound is also indicated by a blue light and the moment the blue light turns

orange, the user must insert the tip into the manifold as far as it will go making sure the tip collar

rests on the manifold shoulder and dispense the liquid in one clean motion and then remove the tip

without aspirating.

1.6.8 Cells and compound additions scheme

1.7 Temperature control

The temperature control ensures temperature stabilization at the recording site throughout the

experiment with a temperature accuracy of ±0.5˚C.

The temperature stabilization interval ranges from 18˚C/64˚F to 40˚C/104˚F.

The temperature control is enabled by the thermoelectric circulator Loop100 (Lauda), connected to

QPC through insulated tubing and a data cable. An ambient temperature sensor on QPC enables

temperature stabilization independent of the ambient temperature.

The thermoelectric circulator heats and cools decalcified water and pumps it through the bed of nails

(BON) inside QPC and thereby stabilizes the recording temperature. The circulator is filled with

decalcified water including an anti-fungal and antibacterial agent through an opening on the top of

the circulator. Instructions on how to fill/empty the thermoelectric circulator is described in section

5.3.

The thermoelectric circulator may display a different temperature on its display compared to the

target temperature indicated in the SSS depending on the ambient temperature and the absolute

temperature indicated. This is expected as the temperature control also takes the ambient

temperature into consideration.

The circulator has an independent power inlet and must be activated in the SSS on the QPC touch

screen to actively stabilize temperature at the recording site. This is explained in a subsequent

section.

Before starting an experiment, the QPC must be activated, and the temperature control dial set to

the desired temperature for 1 hour prior to starting any experiments.

Color

Instruction

Blue

Await orange light without inserting pipette tip

Orange

Dock & dispense, then remove the tip without aspirating

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1.8 Sophion Analyzer software

The Sophion Analyzer software is used for designing and planning experiments and for analyzing

recorded data. It is used to design voltage protocols and whole cell protocols (pressure protocols) as

well as documenting the cell type and ion channel studied. These protocols cannot be modified during

the recording experiment.

It comes pre-installed on the QPC instrument PC or it is downloaded and installed on one or more

office PCs simultaneously, connected to the same network as the instrument in order to access the

Oracle database in the instrument PC.

Both raw data and analyzed data can be displayed. The analysis comprises all elements relevant for

patch clamp data like IV relation, mono- and bi-exponential fit to kinetic parameters as well as

pharmacological quantification.

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2. Startup and shutdown procedures

2.1 Startup

Please see the installation guide (available on: QPatch Compact self-support - Sophion, scan QR

upper right hand corner for access) for details on setting up the instrument.

Before the system is powered up, ensure that the pressure and vacuum inlets are connected and

that the pressure and vacuum levels are within the specified range (see chapter 6 for installation

tests and chapter 7 for instrument specifications).

Make sure the thermoelectric circulator is filled with decalcified water incl. an anti-fungal and anti-

bacterial agent, and that the water circulator is connected to QPC. Inspect the system and ensure

the instrument is clean and dry.

2.1.1 Power up & initialization

Turn on the circulator by pressing the power button on the back of the device. The circulator will

automatically enter standby mode (visible on circulator display).

Then turn on QPatch Compact by pressing the power button on the left side of the instrument, a

white ring will illuminate as long as it is turned on. The touch screen will activate and QPC will start

initializing.

During initialization, the QPC manifold mechanism will move and automatically release the red

transport plate which sits in the QPlate slot during transport. This may take a couple of minutes.

Caution! As the insulated tubing fills with water the low volume circulator container

will drain slightly. Make sure that the container is refilled with decalcified water as

this ensures optimal heating/cooling properties of the temperature control.

2.1.2 Logon

The screening station login screen will appear, and the user must log in with their credentials and

password. New user accounts are set up in the Sophion Analyzer software, using the administrator

logon - consult our Sophion Analyzer user manual for more information.

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Once logged in, the Pre-flight screen will appear and the QPlate icon appears indicating to insert the

QPlate. The light guidance next to the manifold will indicate the direction in which to insert the QPlate.

Figure 3 QPatch Compact preflight screen

2.1.3 Temperature control

Press the temperature control icon in the lower right corner to enter the temperature control settings.

Activate the temperature control by toggling the button and set the desired target temperature for

your next experiment, see Figure 4. Exit the temperature control settings.

We recommend powering up the system 1 hour with the

temperature activated and the target temperature set before

conducting any experiments. This ensures that the

instrument has reached a steady temperature level before

conducting any experiments and will ensure optimal

temperature recording conditions.

Once the QPlate is inserted, the QPlate color and barcode is

shown on the touch screen and the instrument is ready to set

up the experiments.

2.2 Shutdown procedure

On the touch screen monitor click on “Logout” to return to the login screen. Click on “X” in the upper

right corner to shut down the application. It takes a couple of seconds for the touch screen to

respond. You can now turn off the power to the system by clicking the power button on the left-hand

side of the instrument.

Note! To access the database from another laptop through a network connection,

you must leave the instrument turned on.

Figure 4 Temperature control dial

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