Merck Millipore Milliflex User manual

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www.sigmaaldrich.com
Milliex® Quantum Rapid Detection
System
User Guide

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Notice
The information in this document is subject to change without notice
and should not be construed as a commitment by Millipore Corporation.
Millipore Corporation assumes no responsibility for any errors that may
appear in this document. This manual is believed to be complete and
accurate at the time of publication. In no event shall Millipore Corporation
be liable for incidental or consequential damages in connection with or
arising from the use of this manual.

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Contents
Introduction.............................................................................................................4
Chemistry of the Reaction .........................................................................4
Principles of the Procedure ........................................................................ 4
Operator and Equipment Safety..................................................................................5
Specications and Operating Requirements ..................................................................7
System Components .................................................................................................9
The Milliex Quantum Reader and Stand ..................................................... 9
The Milliex Quantum Camera ................................................................... 9
The Milliex Quantum Membrane Transfer Tool ............................................. 9
The Removal Rack.................................................................................. 10
The Milliex PLUS Pump .......................................................................... 10
The Milliex Quantum Consumable Kit ...................................................... 10
Installing the Milliex Quantum Reader ..................................................... 11
Installing the Membrane Transfer Tool ....................................................................... 12
Preparing the Removal Rack..................................................................................... 15
Installing the Camera.............................................................................................. 15
Installing the Software............................................................................ 15
Preparing the Reagent Kit ....................................................................................... 20
Precautions ........................................................................................... 20
Reconstitution........................................................................................ 20
Using the Milliex Labels (optional) ........................................................................... 21
Filtering ................................................................................................................ 22
Incubating............................................................................................................. 22
Staining ................................................................................................................ 22
Installing the Milliex Membrane in the Reader ........................................................... 25
Reading the Membrane............................................................................................ 26
Reading the Membrane Manually .............................................................. 26
Reading the Membrane with the Milliex Quantum Camera .......................... 27
Reincubation ......................................................................................... 32
Storing the Reader ................................................................................. 32
Maintenance .......................................................................................................... 33
Cleaning recommendations...................................................................... 33
Daily Cleaning ....................................................................................... 33
Weekly Servicing.................................................................................... 35
Monthly Servicing................................................................................... 35
Semi-Annual Servicing ............................................................................ 36
Annual Servicing .................................................................................... 36
Troubleshooting...................................................................................................... 39
Spare Parts, Accessories and Services ....................................................................... 40
Standard Warranty ................................................................................................. 41

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Introduction
Milliflex Quantum System is a rapid fluorescence based technology
for fast quantitative detection of contaminants over a broad range of
filterable matrices. This easy to use and simple system can detect viable
microorganisms filtered on a membrane down to one CFU per sample. The
non-destructive method allows the identification, by any method, of any
microorganisms detected during the initial fluorescence reading. Sample
preparation is performed by means of membrane filtration using the
Milliflex system that ensures consistent and reliable results.
Chemistry of the Reaction
The Milliflex Quantum system uses proven technology based on an universal
enzymatic fluorescent staining of viable microorganisms.
After a filtration step with Milliflex sample preparation devices, all
microorganisms retained on the membrane are stained by a fluorescence
viability marker. The reaction consists of an enzymatic cleavage by active
microbial metabolism of a non-fluorescent substrate. Once cleaved by
the cell’s metabolism, the substrate liberates free fluorochrome into the
microorganism cytoplasm that cannot leave the cell due to its hydrophilic
nature.
The signal is thereby naturally amplified by accumulation of the
fluorochrome inside the cells and can be easily detected by exposure to
the excitation wavelength of the fluorescence dye in the Milliflex Quantum
reader and visually counted (or via usage of Milliflex Quantum Camera and
the dedicated software).
Principles of the Procedure
The procedure is easy and requires limited training & skills: sample
contaminants are collected by membrane filtration using the Millipore’s
Milliflex system. After a short incubation time, media cassettes are
removed. Each Milliflex membrane is then transferred onto a cellulose pad
pre-soaked with 2 ml of staining solution and incubated for 30 minutes at
32.5°C ± 2.5 °C. Fluorescent micro-colonies are counted using the Milliflex
Quantum reader or on a PC screen using a dedicated camera and software.
After reading, membranes can be re-incubated on media for later micro-
organisms collection and identification if contamination of the sample was
detected.

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Operator and Equipment Safety
All employees who will operate and/or be near the Milliflex Quantum must comply with the following:
• Read and understand the user guide of the Milliex Quantum before using this Milliex Quantum.
Failure to follow operating instructions could result in user injury or damage to the instrument.
• Read and understand all maintenance instructions in this user guide before performing maintenance
on the pump. Failure to follow instructions could result in user injury or damage to the instrument.
• Any alteration of the pump from factory specication may cause unsafe conditions, and will void the
product warranty.
• Any attempt to use the Milliex Quantum in a manner not specied in this user guide may result in
damage to the instrument, operator injury, and will void the product warranty.
• Do not attempt to open and repair the Milliex Quantum. Service should be performed by trained and
authorized personnel only.
• Place the Milliex Quantum on a clean, at, stable, horizontal surface, away from any source of
excessive heat and close to an easily accessible, properly grounded power supply outlet.
Do not locate the Milliex Quantum under direct lab light, as this could interfere with the reading of
the membrane.
• The Milliex Quantum must be installed and used in a clean and dry area.
• The product is not intended to be used in explosive atmosphere
• Do not expose the Reader , Camera , power supply to liquid. If this happens, immediately switch o
and disconnect the pump from the power outlet, wipe up the liquid
• Use only accessories and replacement parts designed for the pump. Using accessories not designed
for the pump could result in user injury or damage to the instrument.
• When ltrating hazardous liquids, wear and use proper protective clothing and equipment in
accordance with the MSDS for the handling and the disposal of the liquid to be ltrated.
• The system power cord is the main system disconnecting device, and must be easily accessible at all
times.
• To avoid damaging the power cord or the outlet, it is recommended that you pull on the plug and not
on the cord to unplug the system from a wall outlet or an electrical connector.
• Make sure the power cord is not a tripping hazard. Make sure that it is not gripped too tightly at the
plug, the outlet or the point where it is connected to the system.
• Before use, check that the hoses are not unplugged, folded, or damaged.
• In case of skin contact with the ltrated liquid, refer to the safety datasheet of the ltrated liquid for
rst aid measures.
• Dispose the ltrated liquids according to local regulations.
• Before cleaning, shut down the Milliex Quantum and switch o and disconnect the power supply
from power source.
• The power supply must be connected to the earthing system
• The electrical installation must comply with local standards, power supply voltage: 100 - 240 Volt AC,
50-60 Hz.
• Use an electrical surge protector to prevent damage to the system.
• Milliex Quantum is designed for indoor use only.

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Use and storage temperatures
The Milliflex Quantum and/or its accessories must be kept away from any heat source. Do not
expose the system to direct sunlight or to temperatures outside the ranges of values given below.
Use and storage temperatures given below:
System operation System storage Altitude
10 to 40 °C 10 to 40 °C < 2000 m
Maximum relative humidity (storage and operation) 80% at temperatures up to 31 °C, then linear
decrease to 50% RH at 40 °C.

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Specications and Operating Requirements
Dimensions
(nominal)
Component Length cm
(in.)
Width cm
(in.)
Height cm
(in.)
Reader 24.9 (9.8) 14.2 (5.6) 12.5 (4.9)
Reader Stand 30.7 (12.1) 18.0 (7.1) 13.5 (5.3)
Camera 9.7 (3.8) 6.6 (2.6) 11.4 (4.5)
Membrane
Transfer Tool 11.9 (4.7) 8.9 (3.5) 5.0 (2.0)
Removal Rack 25.0 (9.8) 14.0 (5.5) 6.5 (2.6)
Weight (nominal)
Component Weight kg (lb)
Reader 4.4 (9.7)
Reader Stand 1.5 (3.3)
Camera 0.5 (1.1)
Membrane Transfer Tool 1.2 (2.6)
Removal Rack 0.8 (1.8)
Voltage
Catalogue
number
Supply Voltage
Input
Supply Voltage
Output
MXQREAD01 100 V – 240 V
50 Hz - 60 Hz
24V
Direct current
MXQCAM001 Powered by the computer
Power
Catalogue
number
Consumption (mA) Power (Watts)
Nominal Maximal Nominal Maximal
MXQREAD01 670 2080 16 50
MXQCAM001 200 430 1 2.15
Power cord adapter plugs for Europe, North America.
Environmental
Operating
Temperature
15 to 40 °C (59 °F to 104 °F)
Humidity < 90 %
Altitude < 3000 m (9842 ft)
Regulatory
Information
Millipore certies that the Milliex Quantum Reader and the
Milliex Quantum Camera were designed and manufactured in
application of the following European Council directives:
- Electromagnetic compatibility 2004/108/EEC
- ROHS 2002/95/CE

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Materials of
Construction
Component Materials of Construction
Reader
Housing Aluminium sheet 1050 with epoxy
coating
Optical chamber 304L stainless steel
Handle 304L stainless steel
Opical lter Glass
LED protector Transparent polycarbonate
Foot Polyvinyl Chloride (PVC)
Labels Polyester
Screen Protector Polyester
Reader Stand
Stand 304L stainless steel
Foot Styrene butadiene rubber (SBR)
End stops Silicone
Camera
Front plate 304L stainless steel
Housing Aluminium alloy with epoxy coating
Window Glass
Light guide Clear Polymethyl Methacrylate (PMMA)
Membrane Transfer Tool
Housing Black polyoxymethylene (POM)
Body 1.4462 Stainless steel
Lever 1.4462 Stainless steel
Seal Silicone
Screw 316L stainless steel
Removal Rack
Body and adapter Polyphenylsulfone (PPSU)
Pins 316L stainless steel

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System Components
The Milliflex Quantum Reader and Stand
Position
Adjustment
Knob
Viewing Window
Screen Stand
Drawer
Counting Button
On/Off LED
Power
Port
RS232
Port
The Milliflex Quantum Camera
Status LEDs USB Port
Camera Window
The Milliflex Quantum Membrane Transfer Tool
Head
Seal Groove
Ejection Lever

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The Removal Rack
Shuttle
Ejection Tool
Shuttle Handle
Liquid Cassette
Adapter Location
Liquid Media
Cassette Adapter
The Milliflex PLUS Pump
Manual
START
MILLIPORE
Pump Head
Pump Head
Mounting Post
Pump
Control Panel
See the Milliflex PLUS Pump User Guide for details and instructions.
The Milliflex Quantum Consumable Kit
Fluorescence
Reagent
Reconstitution
Reagent
Staining Buffer
1
2
1
2
3
3
Transfer Pipettes
Plastic Storage
Bag
The kit components must be stored at 2 to 8 °C.
48 tests can be performed with one Milliflex Quantum Consumable Kit.

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Installing the Milliex Quantum Reader
The Milliflex Quantum Reader weighs 4.4 kg (9.7 lb) and is designed for use
on a laboratory workbench.
1. Remove all components and their accessories from the boxes. Remove
the reader from the protective bag and place it on a flat, stable,
horizontal surface.
NOTE
Retain all packing materials. If the unit needs to be shipped, it should
be packed in the original materials.
2. Remove the protective film on the window.
3. Place the reader in the stand.
NOTE
The reader stand is not autoclavable.
4. Unscrew the adjustment knob on the stand. Adjust the position of the
reader and tighten the adjustment knob to lock the position.

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1. Connect the end of the power cord to the power outlet on the reader .
LED Power Indicator
2. Connect the power cord to the external power supply. Connect the
external power supply to a power source. The system will start up.
3. The auto test sequence will run and verify that the Milliflex Quantum
Reader is ready to use.

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Installing the Membrane Transfer Tool
NOTE:
The Membrane Transfer Tool should be used in a microbiological safety
cabinet or laminar ow hood.
The Membrane Transfer Tool and seal may be autoclaved for 30
minutes at 121°C or for 10 minutes at 134°C. Do not autoclave the
membrane transfer tool with the seal installed.
1. Install the seal in the groove of the tool as shown below.
2. Ensure that the seal does not twist and is fully seated in the groove.
3. Remove the Milliflex PLUS Pump head from the pump.
4. Place the membrane transfer tool on the pump with the ejection lever
towards the front of the pump.

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1. Press on the membrane transfer tool to lock it in place
2. If the Milliflex PLUS Pump is OFF, turn it on.
3. Run the tare cycle on the pump as described in the Milliflex PLUS Pump
User Guide.
4. The membrane transfer tool is ready to use.

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Preparing the Removal Rack
NOTES
The Removal Rack should be used in a microbiological safety cabinet or
laminar ow hood.
The Removal Rack may be autoclaved for 30 minutes at 121°C or for 10 minutes at
134°C.
The Removal Rack is ready to use and requires no preparation.
Installing the Camera
NOTE
Do not place the reader in the stand if the camera will be installed. Place the
reader on a at, stable surface before installing the camera.
1. Place the camera on the top of the Milliflex Quantum Reader as shown here.
The camera will magnetically lock onto the reader.
Installing the Software
System Requirements
• 2 USB ports.
• Minimum 512 MB of RAM memory, 1 GB recommended.
• Minimum disk space: 500 MB.
• Processor recommended : Intel®Core™2 Duo.
• Minimum screen resolution 1024 x 768 – recommended :1280 x 1024
• Operating system :
Windows XP®32 bits and 64 bits.
Windows Vista®32 bits and 64 bits.
Windows 7®32 bits and 64 bits.
• Microsoft.NET Framework 2.0 SP2 is also recommended. It’s installed
automatically during the installation.
• Adobe Acrobat®Reader 9.0 or higher is needed to display the user guide.
Administrator access to the computer is required to install the Milliflex Quantum
Spot Counter software.

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All screenshots shown are examples and may vary from one computer to another.
1. Switch on the computer.
2. Log on with an administrator account.
3. Insert the installation disc into the CD drive of the PC.
4. Using the file explorer browse to the CD-Rom and double-click the “Setup.
exe” file to start the installation.
5. The installation wizard for the Quantum Spot Counter is displayed. Click Next.
6. Select the language and the destination folder for the software. Click Next.

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1. A new window is displayed, listing the components required to
complete the Milliflex Quantum software installation. Click Install.
2. When the operation is completed, click Next.
3. A message indicating that the software is now ready to be installed is
displayed. Click Install.

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1. A message indicating that the computer must be restarted in order to
complete the installation of the software is displayed. Save and close
all the other applications. Then, select Yes, restart the computer
now and click Finish.
2. After the computer has restarted the QuantumSpotCounter icon is
displayed on the desktop.
NOTE
Do not open the Quantum Spot Counter software before installing the
Milliex Quantum Camera.
3. Connect the small end of the USB cable to the camera.

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1. The other end of the USB cable is equipped with two plugs. Connect
the main plug to a USB port of the PC. Connect the second plug to
another USB port of the PC, if available.
2nd plug
(recommended) Main plug
To the camera
NOTE
The main plug of the camera USB cable must be connected to the
same USB port during installation and use or the camera will not be
recognized by the PC.
2. The PC detects the Milliflex Quantum Camera. Wait until the end of
every components installation.
3. The installation is completed. The software can now be started.

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Preparing the Reagent Kit
Precautions
• Do not use any expired kit component.
• Remove the kit from the refrigerator one hour before starting the
reconstitution.
• Reconstitute under a laminar flow hood or a microbiological safety
cabinet.
• During the seven days shelf life, do not let the staining solution remain
at room temperature for more than four hours per day.
• When removing the staining solution from the refrigerator, wait one
hour before performing a staining.
• Use appropriate personal protective equipment when using the staining
solution.
• Mix the staining solution before performing the first test of the day.
Reconstitution
Reconstitute the staining solution following the procedure on the Staining
Buffer Vial (vial 3) before the first use of the reagent kit. The reconstitution
date may be noted on the Staining Buffer (vial 3) label.
2
1
1
Store the reconstituted Staining Solution at 2 to 8 °C.
48 tests can be performed with the reconstituted Staining Solution. The
approximate number of remaining tests is indicated on the Staining Buffer
(vial 3) label.
The shelf life of the reconstituted Staining Solution is seven days
NOTE
To extend the shelf life, the reconstituted Staining Solution can be
distributed in up to four aliquots. These aliquots can be stored up to
one month in the freezer. Once thawed, the aliquot must be stored at
2 to 8 °C, protected from light. The shelf-life of the thawed aliquot is
seven days.