User Manual
44 Pages
Preview
Page 1
April 2010
TissueLyser II User Manual For disruption of up to 48 or 192 biological samples
Sample & Assay Technologies
QIAGEN Sample and Assay Technologies QIAGEN is the leading provider of innovative sample and assay technologies, enabling the isolation and detection of contents of any biological sample. Our advanced, high-quality products and services ensure success from sample to result. QIAGEN sets standards in:
Purification of DNA, RNA, and proteins
Nucleic acid and protein assays
microRNA research and RNAi
Automation of sample and assay technologies
Our mission is to enable you to achieve outstanding success and breakthroughs. For more information, visit www.qiagen.com.
Contents
Contents 1
2
3
4
5
6
7
Safety Information
5
1.1
Proper use
5
1.2
Electrical safety
6
1.3
Environment
7
1.4
Biological safety
8
1.5
Chemicals
9
1.6
Waste disposal
9
1.7
Mechanical hazards
10
Introduction
11
2.1
About this user manual
11
2.2
Intended use of the TissueLyser II
11
General Description
13
3.1
TissueLyser II principle
13
3.2
Features of the TissueLyser II system
14
3.3
Applications
20
Installation Procedures
21
4.1
Unpacking the TissueLyser II
21
4.2
Installation requirements
21
4.3
Transporting and repacking the TissueLyser II
23
4.4
Installation of a TissueLyser Adapter Set or Grinding Jar Set
24
4.5
Replacing the fuses
25
Operating Procedures
27
5.1
Sample disruption
27
5.2
Storing disruption parameters
28
5.3
Displaying the accumulated operating time
29
5.4
Displaying the software version
29
Maintenance Procedures
30
6.1
Regular maintenance
30
6.2
Regular safety checks
32
Troubleshooting Guide
33
7.1
Error codes displayed by the TissueLyser II
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3
Contents
Appendix A Technical data
35
Environmental conditions
35
Waste Electrical and Electronic Equipment (WEEE)
37
EC declaration of conformity
38
Appendix B Liability clause
4
35
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TissueLyser II User Manual 04/2010
Safety Information
1 Safety Information Before using the TissueLyser II system, it is essential that you read this user manual carefully and pay particular attention to the safety information. The instructions and safety information in the user manual must be followed to ensure safe operation of the TissueLyser II and to maintain the TissueLyser II in a safe condition. The following types of safety information appear throughout this user manual. WARNING
The term WARNING is used to inform you about situations that could result in personal injury to you or other persons. Details about these circumstances are given in a box like this one.
CAUTION
The term CAUTION is used to inform you about situations that could result in damage to the TissueLyser II or other equipment. Details about these circumstances are given in a box like this one.
The advice given in this manual is intended to supplement, not supersede, the normal safety requirements prevailing in the user’s country.
1.1 Proper use WARNING
Risk of personal injury and material damage Improper use of the TissueLyser II may cause personal injuries or damage to the instrument. Do not attempt to move the TissueLyser II during operation. The TissueLyser II should only be operated by qualified personnel who have been appropriately trained. Servicing of the TissueLyser II should only be performed by QIAGEN or an authorized agent (service technicians). Do not carry out any modifications of the instrument. QIAGEN shall not be held responsible for any injuries or damages resulting from nonobservance of the instructions and safety information in this user manual.
The TissueLyser II should only be used for the applications described in the TissueLyser Handbook. The TissueLyser II should only be used in combination
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Safety Information
with accessories approved by QIAGEN (see Section 3.2, page 14). Do not continue to use the TissueLyser II or TissueLyser accessory if it is damaged. Perform the maintenance as described in Section 6. QIAGEN charges for repairs that are required due to incorrect maintenance. CAUTION
Damage to the instrument Use of adapters and plasticware other than those described in this user manual may lead to damage to the clamps of the TissueLyser II.
CAUTION
Damage to the instrument Avoid spilling water or chemicals onto the TissueLyser II. Damage caused by water or chemical spillage will void your warranty.
In case of emergency, switch off the TissueLyser II at the power switch and unplug the power cord from the power outlet.
1.2 Electrical safety WARNING
Electrical hazard Any interruption of the protective conductor (earth/ground lead) inside or outside the TissueLyser II or disconnection of the protective conductor terminal is likely to make the TissueLyser II dangerous. Intentional interruption is prohibited. Lethal voltages inside the TissueLyser II When the TissueLyser II is connected to line power, terminals may be live, and opening the TissueLyser II or removing parts is likely to expose live parts.
To ensure satisfactory and safe operation of the TissueLyser II, follow the advice below:
The line power cord must be connected to a line power outlet that has a protective conductor (earth/ground).
Do not adjust or replace internal parts of the TissueLyser II.
Do not operate the TissueLyser II with any covers or parts removed.
If liquid has spilled inside the TissueLyser II, switch off the TissueLyser II, disconnect it from the power outlet, and contact QIAGEN Technical Services.
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Safety Information
If the TissueLyser II becomes electrically unsafe, prevent other personnel from operating it, and contact QIAGEN Technical Services; the TissueLyser II may be electrically unsafe when:
It or the line power cord appears to be damaged.
It has been stored under unfavorable conditions for a prolonged period.
It has been subjected to severe transport stresses.
If the TissueLyser II is not in use, switch it off at the power switch. If the TissueLyser II will not be used for a prolonged period of time, unplug the power cord from the power outlet.
1.3 Environment Operating conditions WARNING
Explosive atmosphere The TissueLyser II is not designed for use in an explosive atmosphere.
WARNING
Risk of explosion The TissueLyser II is intended for use with reagents and substances supplied with QIAGEN kits. Use of other reagents and substances may lead to fire or explosion.
CAUTION
Damage to the instrument Direct sunlight may bleach parts of the instrument and cause damage to plastic parts. The TissueLyser II must be located out of direct sunlight.
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Safety Information
1.4 Biological safety Specimens and reagents containing materials from humans should be treated as potentially infectious. Use safe laboratory procedures as outlined in publications such as Biosafety in Microbiological and Biomedical Laboratories, HHS (www.cdc.gov/od/ohs/biosfty/biosfty.htm). Samples Samples may contain infectious agents. You should be aware of the health hazard presented by such agents and should use, store, and dispose of such samples according to the required safety regulations. WARNING
Samples containing infectious agents Some samples used with the TissueLyser II may contain infectious agents. Handle such samples with the greatest of care and in accordance with the required safety regulations. Always wear safety glasses, 2 pairs of gloves, and a lab coat. The responsible body (e.g., laboratory manager) must take the necessary precautions to ensure that the surrounding workplace is safe, and that TissueLyser II operators are suitably trained and not exposed to hazardous levels of infectious agents as defined in the applicable Material Safety Data Sheets (MSDSs) or OSHA,* ACGIH,† or COSHH‡ documents. Venting for fumes and disposal of wastes must be in accordance with all national, state, and local health and safety regulations and laws.
* OSHA: Occupational Safety and Health Administration (United States of America). † ACGIH: American Conference of Government Industrial Hygienists (United States of America). ‡ COSHH: Control of Substances Hazardous to Health (United Kingdom).
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Safety Information
1.5 Chemicals WARNING
Hazardous chemicals Some chemicals used with the TissueLyser II may be hazardous or may become hazardous after completion of the sample disruption process. Always wear safety glasses, gloves, and a lab coat. The responsible body (e.g., laboratory manager) must take the necessary precautions to ensure that the surrounding workplace is safe and that TissueLyser II operators are not exposed to hazardous levels of toxic substances (chemical or biological) as defined in the applicable Material Safety Data Sheets (MSDSs) or OSHA,* ACGIH,† or COSHH‡ documents. Venting for fumes and disposal of wastes must be in accordance with all national, state, and local health and safety regulations and laws.
* OSHA: Occupational Safety and Health Administration (United States of America). † ACGIH: American Conference of Government Industrial Hygienists (United States of America). ‡ COSHH: Control of Substances Hazardous to Health (United Kingdom).
Toxic fumes If working with volatile solvents, toxic substances, etc., you must provide an efficient laboratory ventilation system to remove vapors that may be produced. WARNING
Toxic fumes Do not use bleach to clean or disinfect used labware. Bleach in contact with salts from the buffers used can produce toxic fumes.
1.6 Waste disposal Used labware, such as sample tubes, may contain hazardous chemicals or infectious agents from the sample disruption process. Such wastes must be collected and disposed of properly according to local safety regulations. For information on how to dispose of the TissueLyser II, see Appendix A (page 35).
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Safety Information
1.7 Mechanical hazards WARNING
Moving parts Do not leave the TissueLyser II unattended during operation.
To ensure safety, the TissueLyer II will not operate until the transparent hood is closed. WARNING
Moving parts Damage or personal injury can occur if the TissueLyser Adapter Set or Grinding Jar Set is not securely attached. Ensure that both adapters or grinding jars are of similar weight. Do not operate the TissueLyser II with only a single adapter or grinding jar installed.
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Introduction
2 Introduction Thank you for choosing the TissueLyser II system. We are confident it will become an integral part of your laboratory. Before using the TissueLyser II, it is essential you read this user manual carefully and pay particular attention to the safety information. The instructions and safety information in the user manual must be followed to ensure safe operation of the TissueLyser II and to maintain the TissueLyser II in a safe condition.
2.1 About this user manual This user manual provides information about the TissueLyser II in the following sections: 1. 2. 3. 4. 5. 6. 7.
Safety Information Introduction General Description Installation Procedures Operating Procedures Maintenance Procedures Troubleshooting Guide Appendices
The appendices contain the following:
Technical data
Warranty terms
2.2 Intended use of the TissueLyser II The TissueLyser II is intended for molecular biology applications. This product is neither intended for the diagnosis, prevention, or treatment of a disease, nor has it been validated for such use either alone or in combination with other products. Therefore, the performance characteristics of the product for clinical use (i.e., diagnostic, prognostic, therapeutic, or blood banking) are unknown. The TissueLyser II is not designed for continuous operation, but for single 8-hour shifts with 30% operating time. QIAGEN has successfully tested the TissueLyser II with the following adapter sets and grinding beads:
TissueLyser Adapter Set 2 x 96, with one 3 mm diameter tungsten carbide bead or one 5 mm diameter stainless steel bead per collection microtube
TissueLyser Adapter Set 2 x 24, with one 3 mm diameter tungsten carbide bead or one 5 mm or 7 mm diameter stainless steel bead per microcentrifuge tube
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Introduction
Damage to the clamps of the TissueLyser II The clamps of the TissueLyser II are designed for use with the following QIAGEN adapters and consumables:
TissueLyser Adapter Set 2 x 24 (cat. no. 69982) containing standard 2 ml microcentrifuge tubes
TissueLyser Adapter Set 2 x 96 (cat. no. 69984) containing Collection Microtubes (racked) (cat. no. 19560) sealed with Collection Microtube Caps (cat. no. 19566)
QIAGEN adapters together with the recommended tubes form a component that is of an optimal thickness, allowing secure tightening of the clamps of the TissueLyser II without straining the clamping mechanism. Damage to the clamps of the TissueLyser II will occur if other components of reduced thickness are installed. Therefore, the TissueLyser II must not be used with adapters from other suppliers or with QIAGEN adapters containing tubes not recommended by QIAGEN.* Requirements for TissueLyser II users This table covers the general level of competence and training necessary for transportation, installation, use, maintenance, and servicing of the TissueLyser II. Task
Personnel
Training and experience
Transportation
No special requirements
No special requirements
Installation, routine use, and maintenance
Laboratory technicians or equivalent
Appropriately trained and experienced personnel
Servicing
QIAGEN or an authorized agent (service technicians)
* Use of the TissueLyser II with non-QIAGEN adapters has been tested by other suppliers (e.g., Roche recommend use of the TissueLyser II with GS FLX Titanium emPCR Shaker Adapters, LV, which are accessories for the Genome Sequencer FLX Instrument). Use of such adapters has not been tested by QIAGEN.
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General Description
3 General Description The TissueLyser II is a flexible bead mill designed for rapid and efficient disruption of up to 48 or 192 samples at the same time. A wide variety of biological samples can be processed, including human, animal, and plant tissues as well as yeast and bacteria. Each sample is simultaneously disrupted and homogenized by high-speed shaking with a bead in a sealed tube (bead milling).
3.1 TissueLyser II principle Efficient sample disruption is a prerequisite for nucleic acid and protein purification procedures. Incomplete sample disruption can lead to significantly reduced yields and can increase the risk of clogging when using purification columns or magnetic particles. The TissueLyser II thoroughly disrupts and simultaneously homogenizes biological samples in the presence of lysis buffer. Plant tissues can alternatively be disrupted and homogenized in the absence of lysis buffer. Tubes containing sample and bead are placed in a TissueLyser Adapter Set and shaken at a very high speed, causing the samples to be disrupted and homogenized by the beating and grinding effect of the beads. The tubes can be discarded after use, which prevents cross-contamination and provides time savings, as there is no need to clean tubes after disrupting samples. In addition, there is no cross-contamination during sample disruption as each tube is securely sealed with its own lid. The TissueLyser II can also disrupt and homogenize large samples when used in combination with a Grinding Jar Set. Samples are placed in a 10 ml jar containing a grinding ball and shaken at high speed. TissueLyser Adapter Sets, Grinding Jar Sets, and sample tubes can be precooled to minimize degradation of nucleic acids during sample disruption. To find out which procedures require precooling, see the TissueLyser Handbook.
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General Description
3.2 Features of the TissueLyser II system TissueLyser II The TissueLyser II provides high-speed shaking with oscillation frequencies of 3– 30 Hz (180–1800 oscillations per minute). Two clamps at the front of the instrument hold either a TissueLyser Adapter Set or a Grinding Jar Set (Figures 1 and 2). A safety mechanism prevents the instrument from operating until the transparent hood is lowered over the TissueLyser Adapter Set or Grinding Jar Set. Power is switched on and off at the power switch located at the back of the instrument. Oscillation frequency can be adjusted in steps of 0.1 Hz using a keypad (Figure 3) located on top of the instrument. Disruption time, from 10 seconds up to a maximum of 99 minutes, can also be set using the keypad (Figure 3). The disruption parameters for a particular application (i.e., oscillation frequency and disruption time) can be stored in the memory of the TissueLyser II. Up to 9 disruption parameters can be stored.
Figure 1. TissueLyser II (closed). The transparent hood is lowered, and the clamps at the front of the instrument are empty.
Note: The maximum oscillation frequency may drop down to 25 Hz, depending on local fluctuations in the mains voltage.
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General Description
Figure 2. TissueLyser II (open). The transparent hood is raised, and a TissueLyser Adapter Set is installed.
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General Description
D C
B
A
Figure 3. Keypad of the TissueLyser II. A “Memory” display, with “PROG” and “SET” keys for storing disruption parameters. B “Frequency 1/s” display, with “–” and “+” keys for adjusting oscillation frequency. C “Time min/sec” display, with “–” and “+” keys for adjusting disruption time. D “START” and “STOP” keys for starting and stopping operation of the TissueLyser II.
TissueLyser Adapter Set 2 x 24 The TissueLyser Adapter Set 2 x 24 (cat. no. 69982) allows disruption of 48 samples in parallel using standard 2 ml microcentrifuge tubes (e.g., Eppendorf Safe-Lock micro test tubes*). The adapter set contains 2 adapters, with each adapter comprised of a top plate, a bottom plate, and a tube holder for holding 24 microcentrifuge tubes (Figure 4). Sample disruption can be carried out at room temperature or after storing the adapter set at –80°C for at least 2 hours. The adapter set can be cleaned with detergent, microbicides, or up to 96% ethanol. For more information, see the product sheet supplied with the adapter set.
* Use of other tubes in combination with 7 mm diameter beads may result in the beads getting lodged in the lids of the tubes.
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General Description
Figure 4. TissueLyser Adapter Set 2 x 24. Each adapter consists of a top plate, a bottom plate, and a tube holder for holding 24 microcentrifuge tubes (2 ml).
TissueLyser Adapter Set 2 x 96 The TissueLyser Adapter Set 2 x 96 (cat. no. 69984) allows disruption of 192 samples in parallel using Collection Microtubes (racked) and Collection Microtube Caps (cat. nos. 19560 and 19566). The adapter set contains 2 adapters, with each adapter comprised of a top plate and a bottom plate for holding a rack of capped collection microtubes (Figure 5). Sample disruption can be carried out at room temperature or after storing the adapter set at –80°C for at least 2 hours. The adapter set can be cleaned with detergent, microbicides, or up to 96% ethanol. For more information, see the product sheet supplied with the adapter set.
Figure 5. TissueLyser Adapter Set 2 x 96. Each adapter consists of a top plate and a bottom plate for holding a rack of capped collection microtubes.
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General Description
Grinding Jar Set, S. Steel The Grinding Jar Set, S. Steel (cat. no. 69985) allows disruption of 2 large samples in parallel using stainless steel grinding balls (Figure 6). Sample disruption can be carried out at room temperature or after freezing the grinding jars in liquid nitrogen. For more information, see the product sheet supplied with the Grinding Jar Set.
Figure 6. Grinding Jar Set, S. Steel. Each 10 ml jar is made of stainless steel and contains a 20 mm diameter stainless steel grinding ball.
Grinding Jar Set, Teflon® The Grinding Jar Set, Teflon (cat. no. 69986) allows disruption of 2 large samples in parallel using Teflon grinding balls (Figure 7). Sample disruption can be carried out at room temperature. For more information, see the product sheet supplied with the Grinding Jar Set.
Figure 7. Grinding Jar Set, Teflon. Each 10 ml jar is made of Teflon and contains a 20 mm diameter Teflon grinding ball.
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General Description
TissueLyser Single-Bead Dispensers TissueLyser Single-Bead Dispensers dispense individual beads into any sample container (Figure 8). They are available in 2 sizes:
TissueLyser Single-Bead Dispenser, 5 mm (cat. no. 69965): Dispenses beads of 5 mm diameter, and contains a reservoir that can hold approximately 150 beads
TissueLyser Single-Bead Dispenser, 7 mm (cat. no. 69967): Dispenses beads of 7 mm diameter, and contains a reservoir that can hold approximately 45 beads
The dispensers can be cleaned with water or detergent. For more information, see the product sheet supplied with the TissueLyser Single-Bead Dispenser.
Figure 8. TissueLyser Single-Bead Dispenser. The dispenser dispenses individual beads into any sample container.
TissueLyser Bead Dispensers, 96-Well TissueLyser Bead Dispensers, 96-Well dispense 96 beads in parallel into Collection Microtubes (racked) (Figure 9). They are available in 2 sizes:
TissueLyser 3 mm Bead Dispenser, 96-Well (cat. no. 69973): Dispenses beads of 3 mm diameter, and contains a reservoir that can hold approximately 1000 beads
TissueLyser 5 mm Bead Dispenser, 96-Well (cat. no. 69975): Dispenses beads of 5 mm diameter, and contains a reservoir that can hold approximately 300 beads
The dispensers can be cleaned with water or detergent. For more information, see the product sheet supplied with the TissueLyser Bead Dispenser, 96-Well.
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General Description
Figure 9. TissueLyser Bead Dispenser, 96-Well. The dispenser dispenses 96 beads into a rack of collection microtubes.
Beads The following beads for the TissueLyser II are available from QIAGEN:
Stainless Steel Beads, 5 mm (cat. no. 69989)
Stainless Steel Beads, 7 mm (cat. no. 69990)
Tungsten Carbide Beads, 3 mm (cat. no. 69997)
3.3 Applications The TissueLyser II enables disruption and homogenization of:
Human and animal tissues
Human and animal cells
Plant tissues
Yeast
Gram-positive and gram-negative bacteria
For information about specific applications and protocols for sample disruption, see the TissueLyser Handbook.
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