PR1-OM-C Parsortix PR1 User Manual - ANGLE PLC · PR1‐OM‐C Parsortix PR1 User Manual 1 1 Before you start 1.1 What is the Parsortix PR1? The Parsortix system enables the separation

User Manual for Parsortix PR1 Cell Separation System For Research Use Only Not for Clinical or Diagnostic use

Version: PR1-OM-C Parsortix PR1 User Manual Dated: 13 October 2015

© ANGLE Europe 2015

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PR1‐OM‐CParsortixPR1UserManual

©ANGLEEurope2015


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Contents

1 Before you start .......................................................................................... 1

1.1 What is the Parsortix PR1? .................................................................. 1

1.2 Get to know the equipment ................................................................ 1

1.3 Handling the separation cassettes ...................................................... 3

1.4 Process workflow ................................................................................ 5

1.5 Collecting and handling blood samples ............................................... 5

2 How to use the Parsortix PR1 ..................................................................... 6

2.1 Instrument protocols ........................................................................... 6

2.2 Initial checklist ..................................................................................... 6

2.3 Priming................................................................................................. 8

2.4 Blood separation ................................................................................. 9

2.5 Cell harvest ........................................................................................ 11

2.6 Cleaning ............................................................................................. 13

3 In‐cassette immunofluorescence staining ............................................... 14

3.1 Background ........................................................................................ 14

3.2 Procedure .......................................................................................... 14

4 Maintenance ............................................................................................ 16

5 Troubleshooting ....................................................................................... 18

6 Assembly and installation ......................................................................... 19

6.1 Positioning and interference ............................................................. 19

6.2 Unpack the box .................................................................................. 19

6.3 Attach the drip tray ........................................................................... 19

6.4 Apply the non‐slip pads ..................................................................... 20

6.5 Connect the cassette clamp .............................................................. 22

6.6 Connect the reagent tubes ................................................................ 23

6.7 Connect the harvest valve waste tube .............................................. 24

6.8 Start the Parsortix PR1 ...................................................................... 24

6.9 Before first use .................................................................................. 25

7 Technical specifications ............................................................................ 26

8 Health and safety ..................................................................................... 27

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8.1 Potential hazards .............................................................................. 27

8.1.1 Biohazards ................................................................................. 28

8.1.2 Handling reagents and solutions ............................................... 29

8.2 Labelling ............................................................................................ 29

9 Technical support ...................................................................................... 32

9.1 Contact us ......................................................................................... 32

9.2 Consumables and spares .................................................................. 32

9.3 Material compatibility ....................................................................... 34

10 Disposal of electronic equipment ............................................................. 35

11 Certificate of decontamination ................................................................. 36

12 Declaration of conformity ......................................................................... 37

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©ANGLEEurope2015

1PR1‐OM‐CParsortixPR1UserManual

1 Beforeyoustart

1.1 WhatistheParsortixPR1?

The Parsortix system enables the separation and capture of cells present in

blood based on their size and deformability. It comprises an instrument, the

Parsortix PR1, and a series of disposable separation cassettes that can be

selected according to need.

The equipment’s intended use is to filter large, clinically relevant cells from a

liquid sample, typically blood. Examples include circulating tumor cells (CTCs)

and fetal cells.

The key aspects of the system are:

1. The Parsortix PR1: a laboratory benchtop instrument.

2. The separation cassette: a disposable, non‐sterile unit containing a

separation structure comprising a series of steps across which cells are

forced to pass.

3. The separation principle: cells separate on the combined basis of their

size and deformability.

4. Capture of target cells: target cells (e.g. CTCs and fetal trophoblasts)

are captured on a separation “step” inside the cassette.

5. Protocols: the system is operated via a set of ready to use automated

procedures.

1.2 Gettoknowtheequipment

Key components of the Parsortix PR1 (Figure 1.1) are:

1. Screen and control panel

2. Cassette clamp (holding a separation cassette)

3. 500 ml waste container

4. 250 ml buffer reservoir

5. Blood sample tube (Vacutainer or Falcon tube)

6. 15 ml tubes (x6) for staining reagents

WARNING: The Parsortix PR1 is for RESEARCH USE ONLY and not for use in clinical or diagnostic procedures.

WARNING: The device is intended for use by suitably qualified and experienced personnel in laboratory environments.

WARNING: It is essential to read the Health and Safety Information presented in Section 8 before assembling or operating the Parsortix system.

Important information

Older versions of the instrument may be named Parsorter PR1, but there is no difference between instruments with these alternative names, nor in their operation. The name Parsortix PR1 will be used throughout this document.

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2 PR1‐OM‐CParsortixPR1UserManual

7. 50 ml tubes (x2) for cleaning and priming reagents

8. Harvesting valve

9. Harvest waste tube

Figure 1.1 Overview of key components of the Parsortix PR1.

The screen (Figure 1.2) displays information about the system status and

prompts for user input. The four buttons are soft keys that invoke various

functions as displayed in the screen above each button.

Figure 1.2 Control panel showing the main menu on screen.

The main menu, shown in Figure 1.2, allows access to the protocol library. In

order to browse the list of available protocols use the keys for left [<<] and

right [>>] to scroll then press [Run] to activate the selected protocol. When the

system is running, two soft keys will be allocated the functions [Abort] and

[Esc]. Pressing and holding [Abort] will stop the protocol and return to the

main menu. Following an [Abort] action, the instrument must be reset by

running a cleaning process (Section 2.6). Pressing and holding [Esc] will

WARNING: Risk of eye damage. This equipment has several sections of flexible plastic tubing that are not fixed. Handle with care to avoid tubing striking eyes.

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progress the system to the next activity in the current protocol. The key

[System] is generally used for instrument servicing and in cases of operational

problems in consultation with Angle Technical Support. In normal use, it

should not be operated. If the system control is activated by mistake the

screen will show three controls: [USB], [Shutdown] and [Exit]. Press [Exit] to

return to the main menu.

1.3 Handlingtheseparationcassettes

Cassettes are available with different gap sizes. Contact ANGLE for details.

During normal operation of the Parsortix PR1, two cassettes are required: a

separation cassette and a cleaning cassette. They are both a Parsortix GEN3

Separation Cassette but each is assigned to a different process. The separation

cassette is a new cassette requested during the priming process for the

subsequent separation of a blood sample. The cleaning cassette is used during

the intermediate and final cleaning processes (use a GEN3P10 or GEN3D10

cassette for cleaning). The cleaning cassette can be reused. A cleaning cassette

in repeated use should be changed after approximately one month.

The cassette in use is placed inside the cassette clamp. To remove and insert a

new cassette:

1. Hold the clamp upside down to avoid dropping the cassette. Release

the hinged retainer and open the clamp.

2. Remove the cassette already in place. Retain cleaning cassettes for

subsequent processes. Retain separation cassettes for continuing use

(e.g. harvest, staining, analysis, etc.) or dispose of them if no longer

required.

3. Insert the cassette required for the protocol in progress. The cassette

must be inserted in the correct orientation. To achieve this, the

cassette alignment slots should sit on the clamp retaining pins and the

orientation arrows in the cassette must coincide with the inlet and

outlet direction as labelled on the cassette tubing (Figure 1.3).

4. Carefully close the clamp and secure with the hinged retainer.

WARNING: Used cassettes should be handled and disposed of as a bio‐hazard.


The separation cassettes are fragile and must be handled with care. ALWAYS handle the cassette by the edges and avoid applying pressure to its surfaces.

Useful tip

Examining your cassette to find the side with the film will help orient the slots. With the cassette film side up, the slots are beneath the film.

To close the clamp, slightly pull the hinged retainer away and over the clamp.

WARNING: Risk of aerosol. Never attempt to remove a Vacutainer from its mount, or a cassette from its clamp, whilst instrument is in operation.

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Figure 1.3 Positioning of the separation cassette inside the clamp. Notice the arrows showing the cassette alignment slots and the corresponding retaining pins on the clamp.

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1.4 Processworkflow

The operation of the Parsortix PR1 is divided into distinct processes controlled

by software based protocols. The workflow is summarised in Figure 1.4.

Figure 1.4 Parsortix PR1 workflow.

1.5 Collectingandhandlingbloodsamples

Blood for separation in the Parsortix PR1 must be drawn into an EDTA

Vacutainer to inhibit coagulation. It is strongly recommended to collect blood

samples into a Becton Dickinson 10 ml Vacutainer, product number 366643

(US) or 367525 (UK). The Parsortix PR1 is specifically designed to fit this

Vacutainer.

The Vacutainer used for phlebotomy can be attached directly to the

instrument or, alternatively, a fraction of the sample can be separated into a

second Vacutainer. In this case, ensure that the second Vacutainer has been

rinsed with PBS to avoid exposing the blood sample to a higher concentration

of EDTA.

The Parsortix PR1 also accommodates a 50 ml tube for use with larger sample

volumes. The mount fits a specific 50 ml tube (Corning product number

352070).

In order to avoid spillages, a blood tube should only contain up to 80% of its

total volume.

In‐cassette staining Cell harvest

Short clean

Priming

Blood separation

Full clean


Blood drawn into EDTA Vacutainers should be stored at 4°C if it is not to be immediately separated. When stored in this way blood samples can be separated successfully on the Parsortix system for up to 48 hours post draw.

WARNING: Do not mount blood sample tubes made of glass onto the Parsortix PR1 as there is a risk of breakage and injury.

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2 HowtousetheParsortixPR1

2.1 Instrumentprotocols

The series of processes performed in routine operation, summarised in Figure

1.4, are implemented by means of automated protocols embedded in the

Parsortix PR1. The processes and their respective protocols are summarised in

Table 2.1.

Table 2.1 Operation processes of the Parsortix PR1.

Process Description Protocol name Time (approx.)

Priming

Prepares the instrument and a new cassette to receive a blood sample

PX2_P 15 min

Separation A blood sample is placed on the instrument and separated in the cassette

PX2_S99F 2 h for an 8 ml sample

Pre‐harvest flush Flushing the instrument separation flowpath to remove residual blood cells prior to harvest

PX2_CT2 25 min

Harvest Removal of the cells captured in the cassette and collection for downstream analysis

PX2_H 5 min

In‐cassette staining Immune fluorescent staining of captured cells

PX2_S99F_stain4H 6.5 h

Short clean Cleans and resets the instrument

PX2_CT 45 min

Full clean Cleans and resets the instrument including the staining reagent lines

PX2_C 1 h 15 min

2.2 Initialchecklist

Before running a protocol on the Parsortix PR1, it is important to ensure that

the following conditions are fulfilled:

1. The reagents required for normal operation are in place according to

Table 2.2.

2. The Parsortix PR1 must have been cleaned after the last operation.

3. An empty 50 ml Falcon tube and a cleaning cassette are in place.

4. The reagent tubes 1‐6 are empty and the lines are clean.

5. The harvesting valve is turned clockwise

6. The harvest waste tube is empty.


First time here? Go to Section 8 for important Health and Safety advice.


The liquid in the waste reservoir must be less than 400 ml to avoid overfilling and system pressurisation. If necessary, empty the bottle and replace 50 ml Distel before running a protocol.

If the waste reservoir is accidentally overfilled (see section 2.2), abort the protocol and switch off the instrument.

The system does not provide feedback regarding air or incorrect fluids entering the instrument, nor if the waste reservoir is full.

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Table 2.2 Reagents required for normal operation

Reagent Detail Location Volume Comment

Phosphate buffered saline (PBS)

10 mM sodium phosphate / 150 mM sodium chloride, pH 7.3‐7.5

250 ml buffer reservoir

50‐200 ml Other PBS formulations or isotonic saline buffered with other reagents (e.g. Hepes) can be used

Disinfectant: Either Distel or sodium hypochlorite bleach

Undiluted solution

500 ml waste reservoir

50 ml (Empty reservoir when the waste volume reaches 400 ml and replace 50 ml of disinfectant)

50 ml enables a minimum concentration of 10% v/v as bottle fills with waste fluid.

Alkaline cleaning reagent: Either Decon Decomatic or LabKlenz 110

Decon Decomatic, or LabKlenz 110, at 10% v/v in distilled water

50 ml reagent tube connected to line labelled “C”

15‐50 ml

Priming fluid: Ethanol

100% 50 ml reagent tube connected to line labelled “P”

10‐50 ml

Note: Contact ANGLE for a list of branded reagent suppliers.

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2.3 Priming

The priming process prepares the instrument to receive a blood sample. This

process involves filling a new cassette with buffer solution. The priming

process must follow an instrument cleaning process.

To run a priming process:

1. Ensure that the system is ready to run. Refer to the Initial Checklist in

Section 2.2.

2. In the instrument’s main menu screen, select the protocol PX2_P using

the scroll arrows.

3. Press [Run] then [Start].

4. When prompted, insert a new cassette.

5. Press [OK] to continue the priming process.

6. At prompt “Finished P”, press [OK] then [Continue] to return to the

main menu on the screen.

7. This process takes approximately 15 minutes to complete.

Parsortix PR1 workflow

In‐cassette staining

Cell harvest

Short clean

Priming

Blood separation

Full clean

IMPORTANT: A cleaning process must always be run after the last separation, harvest or staining process. If in doubt, the instrument should be cleaned again before running the priming protocol (see the description of the cleaning process in Section 2.6).

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2.4 Bloodseparation

If blood has been stored in a 4°C refrigerator, it should be allowed to

equilibrate to room temperature by placing it on a roller mixer for a minimum

of 20 minutes.

After priming, run a separation process as follows:

1. In the main menu screen, select protocol PX2_S99F and press [Run] then [Start].

2. At prompt “Rinse Vacutainer”

Pull the 50 ml Falcon tube (left in place after the previous cleaning as explained in Section 2.6) off its mount keeping the line inside the tube.

Press [OK] to start the rinse. Collect the rinse fluid inside the Falcon tube.

Fully remove the Falcon tube. Avoid flicking fluid off the tip of the line (see useful tip on sidebar).

3. At prompt “Attach Vacutainer”

Invert the Vacutainer containing the blood sample several times to fully re‐suspend the blood cells, then immediately mount it onto the instrument.

A twisting action will help to push the Vacutainer over the O‐ring and into position.

Secure the Vacutainer by pushing it into the vertical position (Figure 2.1).

Press [OK].

Figure 2.1 Blood sample Vacutainer in the secure vertical position.

4. At prompt “Start?”

Press [OK] to start the separation. This process takes approximately 2 hours for an 8‐10 ml sample.

5. After approximately 30 min, re‐suspend the settled blood cells by tapping the bottom of the Vacutainer while it sits in the inclined position (Figure 2.2).



Cell harvest

Short clean

Priming

Blood separation

Full clean

Useful tip

Use an alcohol damp wipe to avoid splash from the tip of the line while removing the Vacutainer.

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Figure 2.2 Re‐suspending the settled blood cells by tapping the Vacutainer.

Re‐secure the Vacutainer by pushing it into the vertical position.

6. At prompt “Finished S99F”, press [OK] then [Continue] to return to the main menu on the screen.

At the end of the separation process, the blood should have been drawn from

the Vacutainer and passed through the cassette. The cassette will have been

rinsed with buffer. There should be only trace amounts of blood left in the

Vacutainer and the cassette should appear clear of blood to the eye. At this

stage, the target cells will have been captured in the cassette.

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2.5 Cellharvest

The cell harvesting process enables pulses of liquid to be passed through the

cassette in reverse direction. The liquid displaces the captured cells and

pushes them through the harvest line into a collection tube or multiwell plate.

Cell harvesting can only follow directly from a blood separation process.

In order to minimise residual blood cells, it is strongly recommended to

perform a pre‐harvest flush of the instrument. (This process can be left out if

warranted by operational needs, but doing so will considerably increase the

number of residual nucleated blood cells in harvests.) The pre‐harvest flush

procedure is:

1. In the main menu screen, select the protocol PX2_CT2 and press [Run]

then [Start].

2. At prompt “Insert cleaning cassette”

Open the clamp, remove the separation cassette, examine under microscope if intended and keep safe.

Insert the cleaning cassette and press [OK]. 3. At prompt “Empty rgt tubes”

Ensure reagent tubes are empty.

Press [OK] to start the flush process. This process takes

approximately 25 minutes to complete.

4. Depending on experimental need, the cells captured in the cassette

can be examined by microscope at this point. Otherwise, keep in a

safe place until ready to re‐insert into the clamp.

5. At prompt “Finished CT2” press [OK] then [Continue] to return to the


6. Remove the cleaning cassette and reinsert the separation cassette.

The harvested cells can be collected in different collection vessels such as

Eppendorf tubes or multiwell plates. Once the collection vessel is ready, start

the harvesting process:

1. Ensure the harvest waste tube is empty (labelled 9 on Figure 1.1).

2. In the main menu screen, select the protocol PX2_H and press [Run]

then [Start].

3. When prompted, rotate the harvest valve anticlockwise to the position

HAR and press [OK].

4. At prompt “Start”

Remove the harvest line from the harvest waste tube and

clean it with an alcohol soaked wipe.

Place a collection vessel (e.g. an Eppendorf tube) beneath the

harvest line

Press [OK] to start the harvest. A volume of 200 l will flow through the harvest line.



Cell harvest

Short clean

Priming

Blood separation

Full clean

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5. At prompt “Further flush?”

Press [Yes] or [No] depending on the requirements for

downstream analysis. If selected YES, a further 1 ml will be

collected.

6. Place the harvest line back in the harvest waste tube, ensuring that

the tip does not sit in cleaning fluid.

7. On prompt, rotate the harvest valve clockwise to the position “SEP”

and press [OK].

8. At prompt “Finished H” press [OK] then [Continue] to return to the



A further 1 ml flush may result in a higher level of white blood cells in the collection tube along with any captured target cells. For high captured cell purity applications a further flush is not recommended.

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2.6 Cleaning

The cleaning process is used to clean the flow path and the critical

components of the system. At the end of a workflow (which, depending on

experimental need may be the end of a separation, of an in‐cassette staining

process, or of a harvest), it is necessary to clean the instrument to reset the

system leaving it ready for the next process. Two cleaning protocols are

available:

A full cleaning process (PTX2_C) is necessary only after in‐cassette

staining (or after other protocols that may use the reagent tubes) to

clean the whole system including lines 1‐6 used for staining reagents.

When reagent lines are not used, a short cleaning process (PTX2_CT)

can be performed at the end of the harvest or, in cases where cells are

not being harvested, at the end of the separation. This process takes

approximately 45 minutes to complete.

It is also recommended to perform a full clean before priming for experiments

involving in‐cassette staining. The full process takes approximately 1 hour 15

minutes.

To run a cleaning process:

1. In the main menu screen, select the appropriate cleaning protocol

(PX2_C for full clean or PX2_CT for short clean) and press [Run] then

[Start].

2. At prompt “Insert cleaning cassette”

Open the clamp and remove the separation cassette.

Insert a cleaning cassette and press [OK]. 3. At prompt “Empty rgt tubes”

Empty reagent tubes 1‐6 if used for in‐cassette staining. Retain

reagents if necessary.

Press [OK] to start the clean process.

4. At prompt “Finished C” or “Finished CT” press [OK] then [Continue] to

return to the main menu on the screen.

5. Empty the fluids from the harvest waste tube.

6. In the case of a full clean process (PX2_C), discard the reagent tubes 1‐

6, and clean the outside of the lines with an alcohol soaked wipe to

remove residual cleaning fluid. Replace the reagent tubes with new

ones.

7. Remove the vacutainer from its mount and discard safely.

8. Clean the O‐ring and the outside of the line using an alcohol soaked wipe.

9. Place a 50 ml Falcon tube on the mount. This tube collects rinse fluid

in the separation process and can be reused.

10. A twisting action will help to push the Falcon tube over the O‐ring and into position.

11. Secure the Falcon tube by pushing it into the vertical position.


The mount must be free of any visible blood residue. If you find it difficult to remove the blood, use a wipe soaked with a mild, dilute cleaning fluid (e.g. Distel or similar), followed by an alcohol soaked wipe.


In‐cassette staining Harvest

Short clean

Priming

Blood separation

Full clean

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3 In‐cassetteimmunofluorescencestaining

3.1 Background

Blood separation can be followed by immunofluorescence staining of the

captured cells for analysis under a fluorescence microscope.

The harvested cells can be stained in‐cassette using the instrument’s reagent

lines.

The automated protocol follows the procedure described below:

1. Fixative is drawn into the cassette and incubated for 20 min.

2. Permeabilization reagent is drawn into the cassette and incubated for

5 min, followed by two additional draws of permeabilization reagent

and incubation for 5 min.

3. Primary antibodies are drawn into the cassette and incubated for 4

hours, followed by PBS wash.

4. Secondary antibodies and DAPI are subsequently drawn into the

cassette and incubated for 1 hour, followed by PBS wash.

The Parsortix PR1 is designed for end‐user versatility. Antibodies and

concentrations should be tested and optimised by the end user before use.

3.2 Procedure

Specifically, it is strongly recommended that no more than 48 hours have

passed between a Full Clean and the instrument’s priming at the start of the

following process.

To prepare for the in‐cassette staining process:

1. Add the following to the reagent tubes using fresh clean 15 ml Falcon

tubes:

Line 1: 2 ml 4% formaldehyde

Line 2: 2 ml 0.1% Triton X‐100 in PBS

Line 3: 1 ml blocking buffer

Line 4: 1 ml primary antibodies (at optimised concentration in

blocking buffer)

Line 5: 1 ml secondary antibodies (at optimised concentration in

blocking buffer containing 1:5000 DAPI)

INFORMATION: Prior to experiments involving in‐cassette staining, the system must be fully cleaned using the Full Clean protocol PX2_C prior to blood separation, which cleans the separation flow path and the reagent lines.

Useful tip

It is possible to perform the separation and the in‐cassette staining as an integrated process using protocol PX2_S99F_stain4h

Ensure that the staining reagents are in place before starting the combined protocol.



Cell harvest

Short clean

Priming

Blood separation

Full clean

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2. In the main menu screen, select protocol PX2_stain4h and press [Run]

then [Start] to initialise the staining process.

3. At prompt “Finished” press [OK] then [Continue] to return to the main

menu on the screen. The cassette can now be removed and visualised

using fluorescence microscopy and the appropriate channels.

4. After in‐cassette staining, a full cleaning process must be run using

protocol PX2_C (Section 2.6).

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4 Maintenance

Following every separation and harvest or in‐cassette staining:

Run a cleaning process to minimise the risk of cross contamination

between experiments and the build‐up of cellular or plasma residue in

the system’s flowpath.

Check the level of fluid in the waste bottle. If more than 400 ml, empty

the container and add 50 ml Distel disinfectant.

Check the level of fluid in the buffer reservoir and add more if

necessary.

On a daily basis, ensure that the instrument is clean and undamaged:

Check that all the external pipework, unions and other fittings are not

kinked or damaged.

Check that the tubing connecting the instrument to the clamp is not

kinked or blocked.

Check that the waste and buffer reservoir connectors are tight at both

the instrument bulkhead and at the respective reservoir lids.

Check that the O‐rings on the Vacutainer mount are in place, clean and

in good condition. (Replacement O‐rings are available from ANGLE.)

Check that the clamp is clean.

Check that the filters on the buffer and waste reservoirs are in place

and are not contaminated or damaged. Replace if necessary.

Clean the external cover of the instrument with a weak detergent or

70% ethanol or isopropanol followed immediately by a water damp

towel.

If necessary, remove and clean the grey housing surrounding the

Vacutainer.

Contact ANGLE Technical Support if kinks, blocks or other damage are

identified.

On a 3‐monthly basis:

Replace the filters on the buffer and waste reservoirs (see Section 9.2

for part numbers).

Replace the two O‐rings on the Vacutainer mount.

Important information:

Contact ANGLE Technical

Support when damage to

any component of the

instrument is found or

suspected.

Contact details can be

found in Section 9.1.

CAUTION: The routine maintenance plan must be complied with at all times to ensure results are not adversely affected by contamination or sub‐optimal system performance.

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On an annual basis and in conjunction with ANGLE Technical Support:

Replace the internal filters and precision syringe components.

Calibrate and check the integrity of the internal pneumatic system.

Check the integrity of the internal fluidic system.

Replace the moving components in accordance with the instrument

service schedule.

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5 TroubleshootingThe following flowchart describes the recommended actions to address

problems during the separation procedure.

Contact Angle Technical Support if further advice is required.

Figure 5.1 Actions for addressing common problems during the separation procedure.


Biohazard: Clean leaks

safely following local

Health and Safety

procedures. Relevant

advice can be found in

Section 8.1.1.

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6 AssemblyandinstallationCarefully follow the instructions outlined in this Section to assemble and install

the Parsortix PR1.

6.1 Positioningandinterference

The Parsortix PR1 device complies with the emission and immunity

requirements described in the IEC 61326 standard and as set out in the

declaration of conformity in Section 12 of these instructions.

This device complies with Part 15 of the FCC Rules. Operation is subject to the

condition that this device does not cause harmful interference and this device

must accept any interference received, including interference that may cause

undesired operation.

Do not use this device in close proximity to sources of strong electromagnetic

radiation (e.g. unshielded intentional RF sources), as these can interfere with

the proper operation.

6.2 Unpackthebox

Remove all the components from the shipping carton and verify that all the

items listed in the enclosed contents list are present and not damaged. Do not

dispose of the packaging until all the components are removed and accounted

for. Some components (e.g. spare parts) may not be used immediately and

should be stored safely for future use.

Place the instrument in the selected final location on the benchtop. Once

assembled, do not move the equipment whilst the reagent bottles and tubes

are connected as these are not held securely in place and may fall and break.

6.3 Attachthedriptray

The drip tray surrounds the encasing for the blood sample tube and helps

contain spillages or leaks.

Attach the drip tray over the sample mount as shown in Figure 6.1. It is held in

place by a magnetic catch.

WARNING: The electromagnetic environment that this device is to be located within should be evaluated prior to operation of the device.

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Figure 6.1 Positioning of the drip tray.

6.4 Applythenon‐slippads

Adhere the non‐slip pads onto the surface of the reservoir bottles bay on the

left (large pad) and the reagent rack bay on the right (small pad). First, identify

the correct position and then peel the backing paper from the pads and apply

them (Figure 6.2).

Figure 6.2 Placement of the non‐slip pads on the Parsortix PR1: large pad on the left side bay and small pad on the right side bay.

The waste and buffer reservoirs are Duran type glass bottles and are provided

with caps pre‐fitted with tubing, connectors and PTFE filters.

Identify each reservoir bottle and its cap according to the information outlined

in Table 6.1.

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Table 6.1 Identification of bottles and pre‐fitted caps.

Reservoir Bottle volume

Tubing in cap

Inlet tube label

Connection port in the Parsortix PR1

Waste 500 ml Two lines W Left and middle ports Buffer 250 ml Single line B Right port

Figure 6.3 Waste (back) and buffer (front) bottles with pre‐fitted caps.

Attach the caps to the bottles (Figure 6.3) and connect the tubing to the

Parsortix PR1 (Figure 6.4):

Buffer reservoir: carefully remove the red protection cap from the

right connection port on the front of the instrument. Manually screw

the plastic tubing connector into the hole. Place the bottle on the right

of the bottle bay.

Waste reservoir: carefully remove the red protection cap from the left

and middle connection port on the front of the instrument. Manually

screw the plastic tubing connectors into the holes. Place the bottle on

the left of the bottle bay.

Figure 6.4 Connection of the buffer reservoir (left image) and waste bottle (right image).


The connection assembly

comprises a yellow ferrule,

a metal sleeve and a

plastic locking nut. The

yellow ferrule forms a seal

as the plastic nut is

screwed into the

connection port until

resistance is felt then one

further quarter turn .

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6.5 Connectthecassetteclamp

The clamp is the metal case that securely holds the separation cassette during

operation of the Parsortix PR1 (Figure 6.5). It is supplied with its connection

tubes already attached. The inlet and outlet tubes are labelled “I” and “O”,

respectively. The cassette connects to the back of the harvest valve mounted

on the right side of the instrument (Figure 6.6).

Figure 6.5 Cassette clamp (1) connected to the harvest valve (2).

Figure 6.6 Rear view of the harvest valve.

To connect the clamp, carefully remove the red protection covers on the two

unused ports at the back of the harvest valve and connect each line in the

correct port according to the information in Table 6.2.

Table 6.2 Identification of connection ports in the harvest valve.

Line Port number

Position (as viewed from rear of valve)

Inlet “I” 1 Left hand (9 pm)

Outlet “O” 4 Right hand (3 pm)

inlet outlet

©ANGLEEurope2015


6.6 Connectthereagenttubes

The reagent tubes hold the fluids required for priming and cleaning the

system, as well as the reagents for in‐cassette cell staining.

The cluster of tubing lines emerging from the right hand side of the instrument

enables the system to dispense reagents. The lines are tagged with a coloured

label according to the diagram in Figure 6.7. The lines connect to 15 ml or 50

ml plastic tubes held in an 8‐space metal rack placed on the right side bay.

Figure 6.7 Reagent lines identification labels.

To connect the lines, remove the end cap from the line and carefully slide it

through the luer cap of a tube (Figure 6.8). A small amount of force might be

necessary but take care not to kink the line. The line should reach just short of

the bottom of the tube when the cap is screwed on. Refer to the table below

to match the lines with the correct reagent tube. Once the line is connected,

place the tube in the 8‐space metal rack.

Table 6.3 Identification of reagent tubes and lines.

Line Quantity Tube size Use

1‐6 (coloured) 6 15 ml Staining reagents P 1 50 ml Priming fluid C 1 50 ml Cleaning fluid

Figure 6.8 Connection of lines to reagent tubes.

©ANGLEEurope2015


6.7 Connecttheharvestvalvewastetube

The waste fluid produced during the cleaning process is collected in a 50 ml

plastic tube held in a single rack behind the harvest valve.

The harvest valve has an attached line with a free end. To connect this line,

remove the black end cap and carefully slide it through the luer cap of a 50 ml

tube (Figure 6.9). Place the tube on the single rack.

Figure 6.9 Connection of the harvest waste line to the waste tube.

6.8 StarttheParsortixPR1

Connect the power cord to the Parsortix PR1 and to the mains outlet. Turn on

the equipment using the power switch on the left of the instrument. The

internal instrumentation will be heard moving as the start‐up sequence takes

place. The display screen will report the progress before presenting the main

menu with the instruction [Select Protocol] (Figure 6.10). Now the system has

successfully booted and the equipment is ready for use.

Figure 6.10 Control panel showing the main menu on screen.


Do not connect external

equipment to the

instrument, with the

exception of USB drives.

The sockets on the left

hand side are for

manufacturers use only.

©ANGLEEurope2015


6.9 Beforefirstuse

After assembly and before the first use, a new Parsortix PR1 must undergo one

priming and two full cleaning processes to clean, rinse out and reset the

instrument. Refer to Section 2 for instructions on how to run the instrument.

©ANGLEEurope2015


7 Technicalspecifications

Supply voltage 100‐240 V AC 50/60Hz input 24 V DC 4.16 A output

Fuse rating 3.15 A Fast 20 mm

Operating conditions 18°C to 25°C 95‐105kPa

Storage conditions 0°C to 40°C 95‐105kPa

Maximum operating altitude 2000 m

The Parsortix PR1 has been tested in accordance with the following electrical safety standards:

IEC 61010‐1:2010, EN 61010‐1:2010

IEC 61010‐2‐081:2001+A1:2003, EN 61010‐2‐081:2002

UL61010‐1:2012

CAN / CSA C22.2 No. 61010‐1‐12

A Declaration of Conformity for EU markets can be found in Section 12 of this document.

The Parsortix PR1 is certified under certificate number: U8 13 08 85079 001 as compliant with the relevant standards in the US and Canada.

©ANGLEEurope2015


8 HealthandsafetyThe Parsortix cell separation system is designed for use as part of biomedical

research. It must:

Be situated and operated only in facilities with the specialised

infrastructure and general equipment required for biomedical

research or clinical laboratory activity.

Be operated by personnel who have relevant training, knowledge and

experience in biomedical research and/or clinical laboratory

operations, including equipment and materials required to perform

laboratory tasks correctly and safely.

Be used in association with and within existing management and

technical processes concerning correct and safe use of chemicals,

biological materials and other hazards commonly encountered in

biomedical research and clinical operations.

It is not normally necessary to use the equipment within hazard containment

equipment; however, users should consider its use as part of their risk

assessments, taking into account the specific reagents and other materials

being used.

8.1 Potentialhazards

Table 8.1 Risks associated to the operation of the Parsortix PR1.

Risk Description Actions to minimise risk

Biohazards

Exposure to blood and other biohazards can result in infection

Exposure during phlebotomy and sample handling: establish appropriate operating procedures, use containment and personal protective equipment and consider vaccinations. Exposure during equipment operation: adhere to the instructions in Section 2.4 for mounting and removing sample tubes and wear personal protective equipment. Clean spillages as described in this section. Handling of waste: ensure that the waste reservoir is prepared as instructed in Section 2.2 and that the waste material produced is handled and disposed of appropriately. Wear personal protective equipment.

WARNING: The use of the Parsortix cell separation system involves two main potential hazards as described in Table 8.1.

©ANGLEEurope2015


Chemical exposure

Exposure to chemicals that may be caustic, noxious, toxic, volatile or flammable

Establish operation procedures for risk assessment, storage, handling, deployment and disposal of chemical reagents. Wear personal protective equipment.

8.1.1 BiohazardsThe use of the Parsortix cell separation system may involve any or all of the

following biohazards:

blood and its risk of pathogenic infection

other body fluids (e.g. urine) and their risk of pathogenic infection

primary or cultured cells that may be hazardous to health if they enter

a person’s tissues

primary or cultured cells that may themselves be hazardous to health

if they enter a person’s tissues

biochemicals that may be hazardous to health if they contact or enter

a person’s tissues.

Handling bio‐hazardous materials, particularly blood for the Parsortix system,

presents the risk of microbial and viral infection through accidental injury with

contaminated sharps or by exposure to blood, or blood aerosol, without the

use of protective equipment.

The operation of the Parsortix system requires handling blood for two

separate aspects:

phlebotomy and subsequent sample preparation

mounting a blood sample on the instrument during operation

Phlebotomy and subsequent manipulation of blood samples must involve

procedures, containment equipment and protective clothing for protection

against blood and infection. Such measures are likely to form part of

mandatory policies imposed by institutions in which Parsortix equipment will

be used. Vaccination should be considered as a protection measure for all

personnel whose work involves handling or exposure to blood.

The Parsortix PR1 and disposable separation cassettes have been designed to

minimise risk of inadvertent exposure to blood. The design includes a closed

flow path between blood sample tubes and final outflow into waste fluid

receptacles. However, mounting on and removing blood sample tubes from

the equipment involves them being opened and a small risk of blood egress

and exposure from residue present on an instrument component.

Some simple actions negate the minor risk of exposure involved in the

operation of the Parsortix system:

©ANGLEEurope2015


Follow the advice for mounting and removing sample tubes (Section

2.4).

Wearing standard protective equipment including gloves and a face

mask.

In case of a leak during operation (e.g. in the cassette‐clamp

assembly), or if bio‐hazardous material is accidentally spilled on the

instrument, it must first be mopped away using a paper towel or

similar, then cleaned further using 5% Decon Decomatic or equivalent.

Alternatively, if there is particular concern about the biohazard caused

by a spill, after the material is mopped away the area can be

decontaminated and cleaned using 10% Hydrogen Peroxide followed

by 5% Decon Decomatic or equivalent.

Hazardous waste produced through instrument operations must be

handled, stored and disposed of with full regard to appropriate

laboratory safety and environmental regulations.

It is not normally necessary to operate the instrument within a containment

cabinet. However, the equipment can be used within containment equipment

if this is preferred, or if the application or situation in which the equipment is

being deployed is considered to involve particular risks of infection. A

horizontal flow cabinet with an air flow that enters from behind the user, and

that allows open access at the front is particularly convenient, especially if a

microscope is to be used to observe separations.

8.1.2 HandlingreagentsandsolutionsAll chemicals and biochemicals required for use with the equipment are those

in common use in biomedical research. Though several are hazardous, their

safe use is possible through policies and procedures commonly applied in

laboratories for risk assessment, storage, handling, deployment and disposal

of chemicals and biological reagents.

8.2 Labelling

The Parsortix PR1 instrument carries the following hazard, warning and

operational labels:

SYMBOL

MEANING

READ INSTRUCTIONS Read the instruction manual for description of principles of operation and details of potential hazards

BIOHAZARD This instrument contains potential biohazards

©ANGLEEurope2015


CE MARK Indicates compliance with a range of European Directives as set out in the accompanying Certificate of Conformity

MANUFACTURER Name and address of the instrument manufacturer

CAUTION Consult the instructions for use for important warning or cautionary information.

WEEE Do not dispose of in domestic waste.

LOT NUMBER Manufacturer’s lot number for traceability of components

SERIAL NUMBER Manufacturer’s serial number for instrument identification

SINGLE USE ONLY The components must not be used more than once to avoid the risk of cross‐contamination

DO NOT USE IF PACKAGING IS DAMAGED

Do not use the labelled component if the packaging is already open in order to avoid the risk of contamination

KEEP AWAY FROM SUNLIGHT

Parts labelled are sensitive to sunlight and should be stored away from sunlight

FRAGILE Indicates that the contents of a package or container are fragile and should be handled with care

10

CONTAIN SUFFICIENT FOR 10 TESTS

Indicates the number (10 as displayed) of tests (or related consumable items) supplied

DIRECT CURRENT Indicates direct current supply to the equipment.

©ANGLEEurope2015


FUSE Location, value and characteristics of the external system fuse.

THIS WAY UP Preferred packaging orientation for storage and shipping.

KEEP DRY Packaging should be kept dry at all times during storage and shipping.

HARVEST/SEPARATE A switchable valve to select cell separation or cell harvesting mode.

NOTIFIED BODY SYMBOL

The product has been safety tested by TÜV for conformity to IEC 61010‐1 or equivalent standards for markets including EU, USA and Canada.

©ANGLEEurope2015


9 Technicalsupport

9.1 Contactus

Manufacturer and authorised manufacturer’s representative in the EU:.

ANGLE Europe Ltd 3 Frederick Sanger Road The Surrey Research Park Guildford, Surrey GU2 7YD United Kingdom Tel: +44 (0) 1483 685830 Email: eu‐[email protected] www.angleplc.com

Authorised manufacturer’s representative in the USA:

ANGLE North America Inc University City Science Center 3711 Market Street Floor 8 Philadelphia PA 19104 Tel: +1 215 966 6240 Email: us‐[email protected] www.angleplc.com

9.2 Consumablesandspares

The following consumables are available from ANGLE, or in the case of

standard laboratory components, a range of scientific equipment and

consumables providers.

Table 9.1 Consumables and spares

Item Supplier and part number

PR1 user manual and user manual CD Parsortix PR1‐IFU Ten disposable Parsortix 10 μm cell separation cassettes

Parsortix GEN3P10 or Parsortix GEN3D10

Parsortix cell separation cassette clamp assembly with inlet and outlet lines attached, each line with assembly for connection to instrument

Parsortix GEN3Clamp

Two spare nut + ring + ferrule + gasket sets for clamp

Parsortix GEN3Gasket

One flushnut tool (for use in adjustment of clamp fittings)

Parsortix GEN3Flushnut

©ANGLEEurope2015


Two replacement O‐rings for blood sample connection

Parsortix O‐rings

One three port bottle lid with filter and port plug fitted (for buffer reservoir)

Parsortix BuffLid

One three port bottle lid with filter fitted (for waste reservoir)

Parsortix WasteLid

One power supply with UK and US mains leads

Parsortix PSU

Two reagent tube racks

Parsortix Rack

Six 15ml reagent tube caps with ferrules fitted

Parsortix 15mlLid

Three 50ml reagent tube caps with ferrules fitted

Parsortix 50mlLid

Two rubber non‐slip mats Parsortix Mats EDTA blood container / phlebotomy tubes BD 10mL Vacutainer™ Plastic Blood Collection Tubes with K2EDTA

BD 366643 (US, Canada) BD 367525 (Europe, Latin America, Asia Pacific, Australia, New Zealand, Canada)

250 ml Glass laboratory bottle (Buffer reservoir)

Fisher Scientific FB33145 or Camlab Part: 1146888

500 ml Glass laboratory bottle (Waste reservoir)

Fisher Scientific BTF‐730‐090F or Camlab Part: 1146890

15 ml Falcon conical reagent tube BD or Corning 352097 50ml Falcon conical reagent tube BD or Corning 352070 Plastic nut for bulkhead union fittings Upchurch LT‐115 ¼‐28 Super

Flangeless PEEK Short Nut Plastic ring and ferrule for bulkhead union fittings

Upchurch P‐259X Super Flangeless Ferrule

Waste reservoir bottle filter PTFE Disposable Syringe Filter; 0.20 μm; 15 mm dia; nonsterile; Female Luer inlet, male Luer outlet. Cole Parmer WZ‐29550‐08 or equivalent

Buffer reservoir bottle filter Nylon Disposable Syringe Filter; 0.22 μm; 13 mm dia; nonsterile; Female Luer inlet, male Luer outlet. Cole Parmer WZ‐81054‐20 or equivalent

©ANGLEEurope2015


9.3 Materialcompatibility

The Parsortix PR1 has been assessed for compatibility with a range of known

aggressive chemicals and reagents as listed below.

Table 2 List of compatible chemicals and reagents.

Chemical/Reagent

Sodium chloride in aqueous solution to a maximum concentration of 0.9% w/v 100% ethanol Hydrogen peroxide in aqueous solution to a maximum concentration of 10% v/v Triton X100 detergent in aqueous solution to a maximum concentration of 10% v/v Tween 20 detergent in aqueous solution to a maximum concentration of 5% v/v Sodium dodecyl sulphate (sodium lauryl sulphate) detergent in aqueous solution to a maximum concentration of 1% w/v Contrad 100 detergent in aqueous solution to a maximum concentration of 10% v/v Decon 90 detergent in aqueous solution to a maximum concentration of 2% v/v Bleach (sodium hypochlorite) in aqueous solution to a maximum concentration of 10%w/v Coomassie blue dye as commercial ready to use reagent Haematoxylin and Eosin dye combination as commercial ready to use reagent Wright’s reagent dye as commercial ready to use reagent Reagent grade Acetone (typically ≥99.5% purity) Reagent grade Butanol (typically ≥99.5% purity) Anhydrous glacial acetic acid 0.1M sodium hydroxide

For assessment of the instrument compatibility with chemicals and reagents

that are not on this list, please contact ANGLE Technical Support prior to

deployment.

©ANGLEEurope2015


10 DisposalofelectronicequipmentIt is important to understand and follow all laws regarding the safe and proper

disposal of electrical instrumentation.

Disposing of this product correctly will save valuable resources and prevent

any potential negative effects on human health and the environment which

could otherwise arise from inappropriate waste handling. If you are unsure of

your national requirements with respect to disposal please contact your local

authority or ANGLE Technical Support for further information.

The symbol of a crossed‐out wheeled bin on the

product means that the device must not be disposed

via the general waste collection system.

In the European Union, this symbol is required in

accordance with the Waste Electrical and Electronic

Equipment (WEEE) Directive 2012/19/EC of the

European Union on all devices put on the European

market after 13/August/2005.

Please contact an ANGLE Technical Support for appropriate decontamination

information and details on the take back program which will facilitate the

proper collection, treatment, recovery, recycling, and safe disposal of the

device.

©ANGLEEurope2015


11 Certificateofdecontamination

An electronic copy of this certificate is available from ANGLE Technical

Support. ANGLE will not accept returned equipment without a valid RMA

number or a fully completed certificate.

©ANGLEEurope2015


12 Declarationofconformity

©ANGLEEurope2015


DO NOT PRINT THIS PAGE Document Control – Information for Users Document Status: FINAL | Dated: 13 October 2015

Document Revision History

Approved by Signature Date

Dr Shane Booth

(Technical) 29 September 2015

Martin Cooke

(Regulatory)

29 September 2015

Michael O’Brien

(Commercial, EU)

13 October 2015

Peggy Robinson

(Commercial US)

09‐Oct‐2015

Version Issue Date Revision Author

OM - C1 1 28 August 2015 First draft for review Akemi Nogiwa

OM-C2 1 9 September 2015 Revision after circulation for comment

Shane Booth

OM-C3 1 23 September 2015

Revision from SB, MC and AN Akemi Nogiwa

OM-C 1 13 October 2015 Final approved version SCB

Documents

PR1-OM-C Parsortix PR1 User Manual - ANGLE PLC · PR1‐OM‐C Parsortix PR1 User Manual 1 1 Before you start 1.1 What is the Parsortix PR1? The Parsortix system enables the separation