Processor Plus User Manual

Important user information All users must read this entire manual to fully understand the safe use of Processor Plus. Important! Processor Plus is intended for research use only, and should not be used in any clinical or in vitro procedures for diagnostic purposes. Processor Plus should not be used in any clinical or in vitro procedures for diagnostic purposes. Safety notices This manual contains warnings and cautions concerning the safe use of the product. See definitions below. WARNING! The WARNING symbol and notice highlight instructions that must be followed to avoid personal injury. Do not proceed until all stated conditions are clearly understood and met.

CAUTION! The CAUTION notice highlights instructions that must be followed to avoid damage to the product or other equipment. Do not proceed until all stated conditions are met and clearly understood.

Notes Note: A Note is used to indicate information that is important for trouble-free and optimal use of the product. Recycling This symbol indicates that the waste of electrical and electronic equipment must not be disposed as unsorted municipal waste and must be collected separately. Please contact an authorized representative of the manufacturer for information concerning the decommissioning of equipment.

WARNING! This is a Class A product. In a domestic environment, it might cause radio interference, in which case the user might be required to take appropriate measures.

WARNING! All repairs should be done by personnel authorized by GE Healthcare. Do not open any covers or replace parts unless specifically stated in the instructions. CE-certification This product complies with the European directives listed below, by fulfilling corresponding standards. A copy of the Declaration of Conformity is available on request. •

2006/95/EC, Low Voltage Directive

•

2004/108/EC, EMC Directive

The CE logo and corresponding declaration of conformity, is valid for the instrument when it is: –

used as a stand-alone unit, or

–

connected to other CE-marked GE Healthcare instruments, or

–

connected to other products recommended or described in this manual, and

–

used in the same state as it was delivered from GE Healthcare except for alterations described in this manual.

Note: The Declaration of conformity is valid only for systems that are marked with the CE logo:

Contents 1

Introduction to Processor Plus ...9 1.1 1.2 1.3 1.4 1.5

2

Automated western blot processing and gel staining ... 9 Processor Plus base parts ... 9 Optional printing ... 9 Protocol key ... 9 Safety precautions ...10

Instrument components ...13 2.1 Processor Plus components and accessories ...13 2.1.1 Tray support...13 2.1.2 Blot processing trays (two sizes)...13 2.1.3 Blot processing tray lid...14 2.1.4 Staining trays (two sizes)...14 2.1.5 Glass lid for gel staining (two sizes)...14 2.1.6 Coated magnets (4) ...14 2.2 Connecting Processor Plus to a serial printer ...14 2.2.1 To connect Processor Plus to the serial printer:...15

3

Navigating the operating software ...17 3.1 Reviewing protocols ...18 3.2 Reviewing the setup parameters ...19

4

Setting up for blot processing ...21 4.1 Configuring the blot processing tray and pump tubing assembly ..21 4.1.1 Waste tube ...22 4.1.2 Vent line ...22 4.2 Leveling the tray ...23 4.3 Calibrating the pump ...24 4.3.1 What’s happening ...25

5

Operating instructions for blot processing ...27 5.1 Prepare for blot processing ...27 5.1.1 Starting at the beginning of the protocol...28 5.1.2 Starting at a particular step in the protocol...29 5.1.3 Interrupting a protocol...30 5.1.4 Pause...30 5.1.5 STOP...30 5.1.6 Liquid in tray ...31 5.2 Cleaning protocol ...31

6

Editing blot processing protocols ...33 6.1 Editing a protocol name ...33 6.2 Editing step variables ...33

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6.2.1 Port In ...34 6.2.2 Minutes ...34 6.2.3 Multiplier ...34 6.2.4 Hold, Beep and Rock (HBR)...35 6.2.5 Editing when Processor Plus is not running a protocol ...35 6.2.6 Adding or inserting a step ...36 6.2.7 Deleting a step ...36 6.2.8 Editing a protocol in progress...36 6.2.9 To change the time in the current step...36 6.2.10 To change a value in a subsequent step...37 6.2.11 To add one or more steps...37 6.2.12 To delete one or more steps...37 6.2.13 Power outages...37

7

Setup parameters for blot processing ... 39 7.1 7.2 7.3 7.4 7.5 7.6 7.7

8

Setup 1: Manual pump operation ...39 Setup 2: Tray size ...40 Setup 3: Tray level ...40 Setup 4: Display protocol volume ...40 Setup 5: Print protocol ...41 Setup 6: Print volume of protocol ...42 Setup 7: Pump calibration ...42

Setting up for gel staining ... 43 8.1 Configuring the gel staining tray and pump tubing assembly 44 8.2 Leveling the tray ...44 8.3 Calibrating the pump ...45 8.3.1 What's happening ...46

9

Operating instructions for gel staining ... 47 9.1 9.2 9.3 9.4

Prepare for gel staining ...47 Starting at the beginning of the protocol ...48 Starting at a particular step in the protocol ...49 Interrupting a protocol ...49 9.4.1 Pause...49 9.4.2 STOP...50 9.5 Cleaning protocol ...50

10 Editing gel staining protocols ... 51 10.1 Editing a protocol name ...51 10.2 Editing step variables ...51 10.2.1 Port In ...52 10.2.2 Minutes ...52 10.2.3 Port Out ...52 10.2.4 Hold, Beep and Rock ...53

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10.2.5 Editing when Processor Plus is not running a protocol ...53 10.2.6 Adding or inserting a step ...54 10.2.7 Deleting a step...54 10.2.8 Editing a protocol in progress...54 10.2.9 To change the time in the current step...54 10.2.10To change a value in a subsequent step ...54 10.2.11To add one or more steps ...55 10.2.12To delete one or more steps ...55 10.2.13Power outages...55

11 Gel staining setup parameters ...57 11.1 Setup 1: Manual pump operation ...57 11.2 Setup 2: Tray size ...58 11.3 Setup 3: Volume delivery ...58 11.4 Setup 4: Tray level ...58 11.5 Setup 5: Print protocol ...59 11.6 Setup 6: Pump calibration ...59

12 Using the protocol key ...61 12.1 Editing with the protocol key in the slot ...61 12.2 To copy a protocol from the key to Processor Plus ...62 12.3 To copy from Processor Plus to the protocol key ...62 12.4 To edit a protocol on the protocol key ...62

13 Troubleshooting ...63 13.1 Pumping irregularities ...63 13.2 Power Outage ...63 13.2.1 Instrument-based problems ...63 13.2.2 Gel Staining problems ...64 13.2.3 Blot Processing problems ...65

14 Care and maintenance ...67 14.1 Running the cleaning protocol ...67 14.2 Staining trays ...67 14.3 Chemical resistance ...69 14.4 Cleaning the splash guard ...69 14.5 Replacing the tray gasket on the staining tray ...69 14.6 Pump care and maintenance ...70 14.6.1 Calibrating delivery volume ...70 14.6.2 Changing reagent lines...70 14.7 Changing the peristaltic pump tubing ...71

Index ... 93

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Introduction to Processor Plus 1

1

Introduction to Processor Plus 1.1

Automated western blot processing and gel staining

Processor Plus automates fluid delivery and timing steps for both membrane processing and polyacrylamide gel staining.

1.2

Processor Plus base parts

•

the electronic and mechanical parts, encased in a metal chassis. The instrument control panel-an LCD and keypad-is on the front of the unit.

•

a peristaltic pump which works in conjunction with the 10-port valve to transfer fluid from reagent bottles to the tray. Both the pump and the 10-port valve are on the back of the unit.

•

memory on Processor Plus that can hold up to 24 protocols: 10 for membranes and 14 for gels. Each protocol can hold up to 30 steps.

A tray and lid sit on top the base unit to hold either the gels or membranes during staining or blot processing. The blot processing tray option includes a tray support, two Mini trays, one Standard tray and one blot processing lid that fits either tray. The gel staining tray option includes a tray support and a choice of two tray sizes- 125 ml and 250 ml-each with its own lid and tray support.

1.3

Optional printing

You can connect Processor Plus to a serial printer via a 9-pin cable and adaptor (code no. 80-6427-70). See Section 2.2 for instructions. When a serial printer is connected to Processor Plus while a protocol is running, a paper report of each step in a protocol is generated, including: •

the volume of reagents used in each step

•

the time, in minutes, for each step

•

a validation report form, with spaces for sample ID, date, and operator

When Processor Plus is not in use, you can use the Setup menus to print all the steps in a protocol or calculate the volume of each reagent needed.

1.4

Protocol key

The protocol key is a removable device that can store one protocol. You can use the protocol key to record a protocol, then remove the protocol key and store it in a safe place. Use the key to repeatedly run the stored protocol. The protocol on the protocol key does not change unless you overwrite it.

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1 Introduction to Processor Plus 1.5 Safety precautions

For more information on using the protocol key, see section 12.

1.5

Safety precautions

1

Read the user manuals before using the product.

2

This instrument is designed for indoor use only.

3

Do not use the equipment where aerosol sprays are being used, nor where oxygen is being administered.

4

Check that the voltage setting matches the supply voltage on your premises.

5

The instrument must always be used with the protective earth lead of the power cord correctly grounded to earth at the main outlet.

6

To permit sufficient cooling, ensure that the vents at the top, rear and bottom of the instrument are not covered.

7

Keep air openings free from lint, hair, fluff etc.

8

Do not place anything on top of the equipment.

9

Unless specifically instructed to do so by this manual, do not drop or put anything into any opening in the equipment, nor into any hose or coupling.

10 Do not operate the instrument in extreme humidity (above 95%). 11 Keep the instrument dry and clean. Wipe regularly with a soft damp tissue. Let the instrument dry completely before use. 12 Never place waste containers on the top of the instruments. If they become full and overflow, liquid may penetrate the instrument causing a short-circuit. If there is a risk that large volumes of spilt liquid may penetrate the casing of the instruments and come into contact with the electrical components, immediately switch off the system and contact an authorized service technician. 13 Any equipment connected to the instrument should meet the requirements of the EN 61 010-1 or other international safety standards. In Europe, connected equipment must also be CE-labelled. 14 Do not operate the product Hardware or Software in any other way than described in the product user manuals. 15 Do not use any accessories, which are not supplied or recommended by the manufacturer. 16 Do not use the equipment if it is not working properly, nor if it has suffered any damage. Examples of damage include:

10

•

damage to the flexible supply cord or its plug

•

damage caused by dropping the equipment

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Introduction to Processor Plus 1

•

damage caused by dropping the equipment into water or splashing water onto it.

17 Do not let the equipment or its flexible cord come into contact with surfaces, which are too hot to touch.

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1 Introduction to Processor Plus 1.5 Safety precautions

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Instrument components 2

2

Instrument components 2.1

2

1

Processor Plus components and accessories

2

1

2

10 7 3

8 6

4 5

9

Fig 2-1. Processor Plus components and accessories Table 2-1. Instrument components.

No.

Component

Function

1 2

Provides the rocking motion for agitating fluid. Fit into the sockets on the bottom of the tray support.

3 4

Rocker finger Pivot ball assemblies (2) LCD Screen Keypad

5 6 7 8

Protocol key slot Reagent lines 10-port valve Peristaltic pump

9 10 11

Pump tubing Protocol key Spirit level

2.1.1

Tray support

Displays protocol status and program steps. Provides keys to program protocols and control Processor Plus. Accepts the protocol key Connects reagent bottles to 10-port valve. Selects reagent line to connect to peristaltic pump. Pumps fluids from the 10-port valve into the tray and out of the tray to waste or bottle. Carries reagents through peristaltic pump. Stores one protocol in removable memory. Assists in levelling trays.

Fits onto the pivot ball assemblies on top of the metal chassis to hold the tray and its lid.

2.1.2

Blot processing trays (two sizes)

The Mini tray has four chambers, each holds a maximum of 50 ml for a 9 × 9.5 cm membrane. The Standard tray has two chambers, each holds a maximum of 125 ml for a 16 × 16 cm membrane. Both trays are disposable.

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2 Instrument components 2.2 Connecting Processor Plus to a serial printer

2.1.3

Blot processing tray lid

A manifold on top of the blot processing lid delivers reagents to the surface of the membranes. Valves on the manifold control delivery to chambers in the trays.

2.1.4

Staining trays (two sizes)

Made of stainless steel, the trays are inert to most staining solutions. Two sizes are available: 19 × 29 and 29 × 35 cm.

2.1.5

Glass lid for gel staining (two sizes)

Lids prevent spills and minimize exposure to staining reagents while allowing observation of the staining process. Two sizes are available: 19 × 29 and 29 × 35 cm.

2.1.6

Coated magnets (4)

When placed on gel staining tray, magnets prevent gels from sliding over one another and from coming into contact with the splash guard. Placed on the corners of gels with plastic backing, they help keep gels submerged.

2.2

Connecting Processor Plus to a serial printer

An RS232C serial port with a 9-pin male D-sub connector is located on the bottom of Processor Plus (Fig 2-2). A 9-pin cable and an adaptor for the serial printer connection is available in a serial printer cable kit (code no. 80-6427-70). The signal and pin number assignments on Processor Plus serial port are: Pin 2 Pin 3 Pin 5 Other pins

Transmit Receive Ground not connected

The serial printer must have the following settings: Baud rate Data bits Stop bit Start bit Parity Flow control

14

1200 8 1 1 None None

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Instrument components 2

2.2.1

To connect Processor Plus to the serial printer:

WARNING! Always turn the mains power switch off and unplug the power cord before doing any maintenance. 18 Attach the 25-pin male adaptor to the port on the serial printer. 19 Connect one end of the 9-pin cable to the RS232C serial port on Processor Plus and the other end to the adaptor on the serial printer. See Fig 2-2.

RS232C serial port 9-pin cable connector

Fig 2-2. Locating the RS232C serial port.

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2 Instrument components 2.2 Connecting Processor Plus to a serial printer

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Navigating the operating software 3

3

Navigating the operating software The LCD on Processor Plus displays the current status of the instrument. When you turn on the power switch, located on the left side of the instrument, a self-diagnostic cycle runs for approximately one minute. This cycle tests all circuits and moving parts. During the self-diagnostic cycle, the pump starts and stops, the instrument emits a “beep,” and the tray rocks. If any component test fails, the self-diagnostic cycle stops and a message on the screen indicates the source of the fault. Press START to skip to the next test. Report component test failures to your nearest GE Healthcare service representative. Once the diagnostic cycle is successfully completed, the screen displays the name of the last protocol run on Processor Plus. Press STOP to return to the Main menu. The keypad (Fig 3-1) contains three types of keys: Program, Arrow and Protocol. Table 3-1. Keypad keys

Type

Key names

Function

Program

EDIT/SET, ADD, DEL

Arrow Protocol

START, STOP, PAUSE

Edit protocols and the variables in each step of a protocol. Move cursor and change values on the LCD screen. Control protocols and setups.

,

,

,

The Main menu is the entry to all protocols and menus on Processor Plus. Use the keypad to navigate from the Main menu to either blot processing or gel staining protocols (see Fig 3-3). program keys

arrow keys

protocol keys

Fig 3-1. The Keypad.

-> Process blot Stain gel Fig 3-2. The Main menu on the LCD.

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3 Navigating the operating software 3.1 Reviewing protocols

3.1

Reviewing protocols

From the Main menu, choose to see either blot processing protocols or gel staining protocols.

Blot processing protocols

Gel staining protocols

1

-≥ Process blot Stain gel

Process blot -≥ Stain gel

or

START

START STOP

Blot Protocol ECL Plus

1

press

Blot Protocol ECL

(Step 1 in Blot Process)

2

Gel Protocol DNA Silver

2

press

EDIT/SET

STOP

3

1

press press

Gel Protocol DNA Silver

4

STOP

2

STOP

EDIT/SET

(Step 1 in Gel Stain Protocol)

5

press

press

press

press

(Step 2 in Blot Process)

STOP

STOP

(Step 2 in Gel Stain Protocol)

Fig 3-3. Navigating through the protocols.

1

From the Main Menu, press and Stain gel protocols.

or

to switch between the set of Process blot

The arrow on the LCD screen indicates which protocol set is selected.

18

2

Press START to see the first protocol in the selected protocol set. Press STOP to return to the Main menu.

3

Press

4

Press EDIT/SET to see the first step in a selected protocol.

5

Press or to review all the steps in the protocol. Press STOP to return to the list of protocols.

•

For more on using the blot processing protocols, go to section 4.

•

For more on using the gel staining protocols, go to section 8.

or

to review all the protocols in the protocol set.

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Navigating the operating software 3

3.2

Reviewing the setup parameters

Processor Plus has two Setup menus, one for blot processing and one for gel staining (see Fig 3-4). Use the parameters in the Setup menus to: •

select a tray size

•

level the tray

•

set the volume of reagent pumped (for gel staining, only)

•

calibrate the pump

•

print a protocol

•

print or display the total volumes of reagents used in the protocol (blot processing, only)

•

manually operate the pump

Blot processing protocols

Gel staining protocols

1

-≥ Process blot Stain gel

or

ST AR T

Process blot -≥ Stain gel ST AR T

STOP

Blot Protocol ECL Plus

1

2

press press press and hold EDIT/SET

Blot Protocol ECL

(Step 1 in Blot processing)

2

STOP

3 4

STOP

Gel Protocol DNA Silver

1

press press

Gel Protocol DNA Silver

press and hold EDIT/SET

2

STOP

(Step 1 in Gel staining)

5

press

press

press

press

(Step 2 in Blot processing)

STOP

STOP

(Step 2 in Gel staining)

Fig 3-4. Navigating to the Setup menus.

1

From the Main Menu, press or to switch between the set of Process blot and Stain gel protocols. The arrow on the LCD screen indicates which protocol set is selected.

2

Press START to see the first protocol in the selected protocol set. Once you have selected the appropriate protocol set, you can go to the Setup menu from any protocol.

3

Press and hold EDIT/SET until the Setup menu appears on the LCD.

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3 Navigating the operating software 3.2 Reviewing the setup parameters

4

Press

or

to review the Setup parameters for the selected protocol set.

5

Press STOP to return to the protocols.

Note: You cannot use the Setup menus when Processor Plus is running a protocol.

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Setting up for blot processing 4

4

Setting up for blot processing Processor Plus allocates memory for ten blot processing protocols, five of these are pre-programmed (Table 4-1). See section 6 for directions on editing protocols. Processor Plus has two trays for blot processing (Table 4-2). One lid fits either tray. Table 4-1. Pre-programmed blot processing protocols

Protocol name

Applications

1 ECL™ Plus 2 ECL 3 Standard 4 Enhanced

ECL Plus Detection Kit ECL Detection Kit Immunodetection with 1º & 2º antibodies Immunodetection with 1º, 2º antibodies and avidin compounds Rinses reagents out of tubing, valves and ports

5 Clean

Table 4-2. Tray Sizes for blot processing

Tray

Number of membranes

Size of membranes (cm)

Reagent capacity (ml /chamber)

Mini Standard

1–4 1 to 2

9 × 9.5 16 × 16

10–50 25–125

4.1

Configuring the blot processing tray and pump tubing assembly

The lid assembly (Fig 4-1) on the membrane processing tray has: •

an inlet manifold in the centre

•

a drain tube at the rear corner

•

four valves for controlling reagent delivery to chambers lid manifold

tray drain tube

inlet manifold tube

Fig 4-1. The membrane processing lid.

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4 Setting up for blot processing 4.1 Configuring the blot processing tray and pump tubing assembly

inner peristaltic pump tube

outer peristaltic pump tube waste tube Fig 4-2. Configuration of membrane processing tubes and tray.

The tubes from the lid assembly connect to the peristaltic pump. The tube from the inlet manifold connects to the inner peristaltic pump. The drain tube in the corner of the lid connects to the outer peristaltic pump. See Fig 4-2. The Quick-Fit couplings on the peristaltic pump tubing have different dimensions, keyed to the appropriate tubing connections on the trays. When the appropriate male and female couplings are pressed together, you can hear a click as they connect.

4.1.1

Waste tube

Place the end of the waste tube into the top of a large waste bottle or suspend it over the edge of the sink. A vented waste bottle cap (38-430), provided with blot processing trays, fits a narrow-mouth Nalgene™ bottle. If you use a waste bottle that does not accept this cap, tape the end of the waste tube to the top of the waste container. Note: Do not let the end of the waste tube become submerged in liquid.

4.1.2

Vent line

IMPORTANT! For membrane processing, reagent line 0 (zero) is reserved for air venting. Do not use reagent line 0 to deliver reagents. The presence of detergent in some reagents creates bubbles. As a result, small amounts of reagent may get into the vent line, but this does not affect membrane processing quality. However, spray from these reagent drops escapes out the vent line when Processor Plus is operating. Suspend the end of the vent line over a sink or large beaker to contain escaping spray. After configuring the tubing and tray, level the tray and calibrate the pump.

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