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BioNetwork Bioprocessing Center Technology Enterprise Center (TEC)

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Presentation on theme: "BioNetwork Bioprocessing Center Technology Enterprise Center (TEC)"— Presentation transcript:

1 BioNetwork Bioprocessing Center Technology Enterprise Center (TEC)
1800G North Greene Street Technology Enterprise Center (TEC) Greenville, NC 27834 fax

2 Staff: William T. (Bill) Cooper, Manager
Greg Smith, PhD, Curriculum Coordinator Vallere Shelton, Administrative Assistant

3 What is the Bioprocessing Center?
Pitt Community College has established a Bioprocessing Center in conjunction with the statewide BioNetwork initiative. It is one of six specialized biotechnology centers developed by the North Carolina Community College System. This Center is a resource center created to promote biotechnology workforce development.  The ultimate goal is to facilitate the growth of biotechnology firms in North Carolina and the surrounding region. Start-up funding for these centers was provided by Golden LEAF.

4 Why Biotechnology? Biotechnology is the fastest growing sector of the economy in the State of North Carolina, a state where traditional jobs, such as textiles and tobacco, are by and large extinct. Biotechnology provides products made from living cells that are useful to all of us. Biotechnology is the wave of the future as scientific and technological discoveries are made daily. Biotechnology jobs pay well.

5 Ranking of U.S. States in the
Number of Biotechnology Jobs

6 State Impact:

7 State Impact:

8 Growth in Protein Technology:

9 How does Industrial Systems fit in?
Because the nature of products created through biotechnology is so valuable to industrial interests, down-time can cost companies millions of dollars in lost profits. Machinery that keeps this industry running needs to be serviced, calibrated, maintained, and cleaned on a regular, expeditious basis. Technicians trained in the culture of biotechnology are more valuable to a company than those that are not. They understand the needs of the industry and have the specialized training to keep the down-time to a minimum. Additionally, they have the expertise to be able to service the equipment without causing undue contamination to the product and the environment.

10 What is Contamination? The introduction of an unwanted substance to a particular process or environment, either intentionally or accidentally; that which has been introduced to a controlled environment. Types of contaminants: Water Soil Air Radiological

11 What is Contamination? Sources of contamination:
Water (Pseudomonas Aeruginosa) Soil (Bacillus Subtilis) Air (any aerosolized form of contamination) Radiological (any unwanted electromagnetic radiation)

12 What is Contamination? Prevention of contamination:
Water (filtration or irradiation) Soil (sanitization and/or disinfection) Air (high efficiency particulate air filters- ultrafiltration) Radiological (shielding or non-use)

13 What is Contamination? Particles are measured in microns. If you were to slice a 1-inch cube into 25,400 equal size pieces, each slice would be 1 micron thick (so thin that you could not see it). Since cleanroom facilities contain invisible particles that can cause defects, we need to have a way to determine their sizes and possible sources.

14 What is Contamination? These particles cannot be seen with the naked eye and yet may cause serious loss to our client and to their ability to manufacture devices without defects. We measure the amount of particles in the air with a device known as a particle counter. This helps us to determine when to perform or restrict certain activities.

15 What is Contamination? To combat the tremendous amount of particulate released by the human body, cleanroom personnel are asked to wear special garments that are restrictive and may be uncomfortable if the working conditions (temperature, humidity) are not carefully controlled.

16 What is Contamination? Typical Pollutant Sizes That Cause Down Time
Smoke .01 to 1.0 Microns Bacteria .25 to 10 Microns Human Skin Flakes 0.4 to 10 Microns Household Dust 0.5 to 100 Microns Mold 2.0 to 20 Microns Human Hair 70 to 100 Microns Pollen 5 to 100 Microns

17 What is Contamination? People are dressed in booties, gloves, helmets, hoods and special facemasks that are not part of a normal experience. You are also asked to move slowly, to clean up dirt you cannot see, and to follow practices and procedures that may not immediately make sense. The approach to cleanrooms is designed in steps so as to ensure that every effort has been made not to introduce contaminants into the cleanroom.

18 What is Contamination? "Humans are walking, breathing, waste pits from a microbiological perspective and there is no indication we are going to evolve into anything cleaner." Human dust combined with sebum will tenaciously adhere to and contaminate surfaces, creating contamination problems that are simultaneously bacterial, chemical and particulate.

19 What is Contamination? There is an additional complicating factor - normal human activity. Humans breathe, sneeze, cough, talk and move around. Body and breath temperatures cause heat turbulence in addition to air turbulence.

20 What is Contamination? People slough off huge numbers of particles a half micron and larger and these are propelled around by air and body movement. Some examples of how many 0.5 micron and larger particles can be released by simple movement are shown below. Vast amounts of contamination need to be controlled in a cleanroom environment.

21 What is Contamination? Activity 0.3µm Particles Released Seated
100,000 particles per minute Head, arm, neck, and leg motion 500,000 particles per minute All of the above with foot motion 1,000,000 particles per minute Standing to sitting position 2,500,000 particles per minute Moving 2 miles per hour 5 million particles per minute Moving 3.5 miles per hour 7.5 million particles per minute Moving 5 miles per hour 10 million particles per minute

22 What is Contamination? Humans also release elemental chemicals that can cause contamination: * Spittle (saliva): potassium, chloride, phosphorus, magnesium, and sodium * Dandruff: calcium, chloride, carbon, and nitrogen * Perspiration: sodium, potassium, chloride, sulfur, aluminum, carbon, and nitrogen * Fingerprints: sodium, potassium, chloride, and phosphorus Consequently, a critical component of cleanroom management is strict adherence to protocol. Cosmetics are prohibited because in addition to their gross particle generation, cosmetics release iron, aluminum, silicone, carbon, titanium, magnesium, potassium, sulfur, and calcium.

23 What is Contamination Control?
Just as humans are the greatest potential contamination risk, they are also the greatest resource for contamination control. A thorough, comprehensive training program detailing all aspects of cleanroom management will empower the cleanroom operators to control the degree of contamination during the production process.

24 What is Contamination Control?
Just as all operators must be thoroughly trained in occupational skills, knowledge of the product, and all aspects of the production process, they must also be trained in the unique practices and behaviors required for working in the cleanroom environment. Testing and certification of cleanroom operators and subsequent observation of operators after training assures that the training has been implemented and is effective. Training is mandatory for all personnel (including Management), regardless of frequency of entry or job classification.

25 What is Contamination Control?
Personal health and hygiene begins at home with daily bathing or showering, shaving, brushing of teeth and hair, and application of silicone-free skin moisturizers to reduce skin flakes. Make-up, hair gels, hair sprays, aromatic after-shave lotions or body lotions are not cleanroom-compatible. While at work, all employees must wash hands after eating and/or using the toilet. Cleanroom-compatible hand cream may be applied prior to gowning.

26 What is Contamination Control?
As mentioned earlier, any activity by the cleanroom operator generates millions of viable and non-viable particles. Consequently, it is imperative to limit talking and actions in the cleanroom to only those required for the manufacture of the product. Running, horseplay, and other non-professional activities are not permitted.

27 What is Contamination Control?
Other behavioral requirements include, but are not limited to, the following: Smoking is not allowed inside the manufacturing facility including all cleanroom areas. Smokers release particles for at least one-half hour after smoking one cigarette.

28 What is Contamination Control?
Nothing is allowed inside the cleanroom complex which is not required in the cleanroom manufacturing process. This includes personal items such as jewelry or keys, cosmetics, tobacco or matches in any form, and food or drink in any form. Hair may not be combed in the cleanroom gowning area.

29 What is Contamination Control?
Only cleanroom compatible ball-point pens are allowed inside the cleanroom for recording data on cleanroom compatible paper and clipboards. While working in the cleanroom, mannerisms such as scratching or hand-rubbing. Cleanroom personnel may not access the inside of the cleanroom uniform.

30 What is Contamination Control?
The use of facial tissues is prohibited in the cleanroom. If one must use a cleanroom compatible non-linting tissue, it must be used only in the gowning area and disposed appropriately in waste receptacle. All doors must remain closed when not entering or exiting. Emergency doors may be alarmed with a visual and audible alarm to enforce compliance.

31 What is Contamination Control?
An integral piece of the cleanroom management program is cleaning of the cleanroom. It is important for all personnel who may work in a cleanroom to understand the critical aspects of maintaining this clean environment.

32 What is Contamination Control?
Below is a partial list of some of the commonly known contaminants that can cause problems in some cleanroom environments. It has been found that many of these contaminants are generated from five basic sources. The facilities, people, tools, fluids and the product being manufactured can all contribute to contamination.

33 What is Contamination Control?
Facilities Walls, floors and ceilings Paint, caulk and coatings Construction material (sheet rock, wood, saw dust etc.) Air conditioning debris Room air and vapors Spills and leaks Rust

34 What is Contamination Control?
People Skin flakes and oil Perspiration Cosmetics and perfume Spit or saliva Coughing or sneezing Clothing debris (lint, fibers etc.) Hair Gum, Cough Drops Food or drink

35 What is Contamination Control?
Tool Generated Friction and wear particles Lubricants and emissions Vibrations Brooms, mops and dusters Paper Cardboard Duct tape Permanent Markers Non-clean room pens

36 What is Contamination Control?
Fluids Particulates floating in air Bacteria, organics and moisture Floor finishes or coatings Thinners or solvents Cleaning chemicals Plasticizers (outgases) Deionized water

37 What is Contamination Control?
Product generated Silicon chips Quartz flakes Cleanroom debris Aluminum particles Packing material Aerosols and smoke

38 Airborne Particulate Cleanliness Classes (by cubic meter)
Monitoring Contamination? Airborne Particulate Cleanliness Classes (by cubic meter) CLASS Number of Particles per Cubic Meter by Micrometer Size 0.1 um 0.2 um 0.3 um 0.5 um 1 um 5 um ISO 1 10 2 ISO 2 100 24 4 ISO 3 1,000 237 102 35 8 ISO 4 10,000 2,370 1,020 352 83 ISO 5 100,000 23,700 10,200 3,520 832 29 ISO 6 1,000,000 237,000 102,000 35,200 8,320 293 ISO 7 352,000 83,200 2,930 ISO 8 3,520,000 832,000 29,300 ISO 9 35,200,000 8,320,000 293,000

39 Colony forming units per volume
Monitoring Contamination? Air Monitoring Class Areas Colony forming units per volume Frequency Class 100 and Class 1000 Critical processing areas for product and container-closures <0.1 cfu per ft3 of air or 3 cfu per m3 Each Shift Class 10,000 Less critical processing areas for product and container-closures <0.5 cfu per ft3 of air or 20 cfu per m3 Daily Class 100,000 Controlled Support Areas 2.5 cfu per ft3 of air or 100 cfu per m3 Twice/week (product/component contact areas) and once/week (other support areas)

40 Monitoring Contamination?
Surface Monitoring Class 100 and Class 1000 Critical processing areas for product and container-closures 3 per 2 inch square ( square cm.) RODAC Plate Each Shift Class 10,000 Less critical processing areas for product and container-closures -5 per 2 inch square (25-30 square cm.) RODAC Plate -10 per 2 inch square (25-30 square cm.) RODAC Plate for the floor Daily Class 100,000 Controlled Support Areas Not Applicable

41 Monitoring Contamination?
Personnel Monitoring Class 100 and Class 1000 Critical processing areas for product and container-closures -3 Gloves (25 – 30 square cm.) -5 Gloves (25 – 30 square cm.) Each Shift Class 10,000 Less critical processing areas for product and container-closures -10 Gloves (25 – 30 square cm.) -20 Gloves (25 – 30 square cm.) Daily Class 100,000 Controlled Support Areas Not Applicable

42 Safety in Controlled Environments:
Cleanroom safety is as important (if not more) than any other facility having laboratories. Workers in these critical environments must be conscientious about not only their own safety, but the safety of the people benefiting from the products produced in the environment.

43 Safety in Controlled Environments:
Everything one learns about safety in a general safety course is applicable in cleanrooms and the responses would be the same. However, in cleanrooms the product is treated with much more respect and the way one behaves is more critical than anywhere else.

44 Safety in Controlled Environments:
Exposure to chemicals and biologics is a risk in cleanrooms just as it is in any laboratory setting, but less likely to occur in controlled environments because of the likelihood of operations ceasing when maintenance or calibration procedures are being performed.

45 Design Considerations and Operation:
The method most easily understood and universally applied is the one suggested by the Federal Standard 209E in which the number of particles equal to or greater than 0.5 microns measured in a cubic foot of air designates the class number. For example, a class 100,000 cleanroom limits the concentration of airborne particles equal to or greater than 0.5 microns to 100,000 particles in a cubic foot of air.

46 Design Considerations and Operation:
Cleanrooms have evolved into two major types which are differentiated by their method of ventilation - turbulent airflow and laminar airflow cleanrooms. The general method of ventilation used in turbulent airflow cleanrooms is similar to that found in buildings such as offices, schools, malls, manufacturing plants, auditoriums, shops, etc. The air is supplied by an air conditioning system through diffusers in the ceiling.

47 Design Considerations and Operation:
A cleanroom differs from an ordinary ventilated room in three ways: increased air supply the use of high efficiency filters room pressurization

48 Design Considerations and Operation:
Increased air supply: The increased air supply is an important aspect of particle control. A typical turbulent airflow cleanroom would have at least 10 air changes per hour and likely have between 20 and 60. This additional air supply is mainly provided to dilute to an acceptable concentration the contamination produced in the room.

49 Design Considerations and Operation:
High efficiency filters: High efficiency filters are used to filter the supply air into a cleanroom to ensure the removal of small particles. The high efficiency filters used in cleanrooms are installed at the point of air discharge into the room.

50 Design Considerations and Operation:
Room Pressurization: Room pressurization is mainly provided to ensure that untreated air does not pass from dirtier adjacent areas into the cleanroom. The cleanroom is positively pressurized with respect to these dirtier areas. This is done by extracting less air from the room than is supplied to it.

51 Design Considerations and Operation:
Laminar airflow is used when low airborne concentrations of particles or bacteria are required. This airflow pattern is in one direction, usually horizontal or vertical at a uniform speed of between 60 to 90 ft/min. and throughout the entire space. The air velocity is sufficient to remove relatively large particles before they settle onto surfaces. Any contaminant released into the air can therefore be immediately removed by this laminar flow of air, whereas the turbulent airflow ventilated system relies on mixing and dilution to remove contamination.

52 Design Considerations and Operation:
Because airflow is such an important aspect of particle control, the design of a cleanroom requires careful consideration of air motion and airflow patterns. Depending on the degree of cleanliness required, it is common for air systems to deliver considerably more air than would be needed solely to meet temperature and humidity design.

53 Design Considerations and Operation:
Airborne particles can be organic or inorganic. Most contamination control problems concern the total contamination within the air. Particles of different sizes behave differently as air moves through a room. For example, in an eight-foot high room, a particle in the 50 micron range might take 60 seconds to settle, while a 1 micron particle might take 15 hours to settle.

54 Design Considerations and Operation:
Before any methods of contamination control of airborne particles can be applied, a decision must be made as to how critical this particulate matter is to the process or product. The quantity of the particles of a given size that might be present within the area must be considered. The source of the contamination is divided into external sources and internal sources.

55 Design Considerations and Operation:
For any given space, there exists the external influence of gross atmospheric contamination. These sources include air pollution in general and dust storms. External contamination is brought in primarily through the air conditioning system. Also, external contamination can infiltrate through building doors, windows and cracks. The external contamination is controlled primarily by the type of filtration used and space pressurization.

56 Design Considerations and Operation:
People and the production process are some of the greatest sources of internal contamination. People in the workspace generate particles in the form of skin flakes, lint, cosmetics, and respiratory emissions. Industry generates particles from combustion processes, chemical vapors, soldering fumes, and cleaning agents.

57 Design Considerations and Operation:
Other sources of internal contamination are generated through the activity of manufacturing equipment. Although airflow design is critical, it alone does not guarantee that cleanroom conditions will be met. Construction finishes, personnel and garments, materials and equipment, and building entrances and exits are other sources of particulate contamination that must also be controlled. Room construction and material finishes are an important part of cleanroom design. Room construction is important to provide an enclosure that will house a process to exclude outside contaminants and that the material finishes will not contribute to particle generation in the space.

58 Design Considerations and Operation:
Walls, floors, ceiling tiles, lighting fixtures, doors, and windows are construction materials that must be carefully selected to meet cleanroom standards. People must wear garments to minimize the release of particles into the space.

59 Design Considerations and Operation:
The types of garments depend on the level of cleanliness required by a process. Smocks, coveralls, gloves, and head and shoe covers are clothing accessories commonly used in clean spaces. Materials and equipment must be cleaned before entering the cleanroom. Room entrances such as air locks and pass trough’s are used to maintain pressure differentials and reduce contaminants. Also, air showers are used to remove contaminants from personnel before entering the clean space.

60 Design Considerations and Operation:
The ability of a filter to remove particles from the air is reflected by its efficiency rating. The American Society of Heating, Refrigerating, and Air Conditioning Engineers (ASHRAE) have developed a standard for measuring filter effectiveness. The standard describes test procedures to classify filters in terms of arrestance and efficiency.

61 Design Considerations and Operation:
Arrestance is the amount of dust removed by the filter, usually represented as a percentage. Since large particles make up most of the weight in an air sample, a filter could remove a fairly high percentage of those particles while having no effect on the numerous small particles in the sample. Thus, filters with an arrestance of 90 percent have little application in cleanrooms.

62 Design Considerations and Operation:
Efficiency measures the ability of the filter to remove the fine particles. ASHRAE efficiencies of between 10 percent and 40 percent should remove 20 percent to 40 percent of the 1 micron particles in the air, but hardly any of the 0.3 to 0.5-micron particles. ASHRAE efficiencies of 80 percent to 95 percent can remove 50 percent to 70 percent of the 0.3-micron particles.

63 Design Considerations and Operation:
A HEPA filter, i.e., high efficiency particulate air filter, is defined by its particle removal efficiency and its airflow rate. A HEPA filter is rated by its efficiency in removing small particles from air and has a minimum efficiency of percent. These high efficiency filters are usually designed to remove particles of 0.3 microns and larger.

64 Design Considerations and Operation:
An ULPA filter, i.e., ultra low penetration air, is a filter that has efficiencies higher than those of a standard HEPA filter. An ULPA filter will have efficiency greater than 99.99 percent. These filters are constructed and will function the same way as a HEPA filters. They differ in that the filter medium that is used has a higher proportion of smaller fibers and is hence more efficient.

65 Design Considerations and Operation:
Most cleanrooms require year-round cooling as a result of the fan energy associated with high cleanroom airflow as well as the heat generated by the process, people, and lighting within the facility. Temperature control is required to provide stable conditions for materials, instruments, and personnel comfort. Human comfort requirements typically call for temperatures in the range of 72F to 75F, since workers frequently wear cleanroom garments over street clothes.

66 Design Considerations and Operation:
Humidity control is necessary to prevent corrosion, condensation on work surfaces, eliminate static electricity, and provide personnel comfort. The human comfort zone is generally in the range of 30 percent to 70 percent relative humidity.

67 Design Considerations and Operation:
A cleanroom facility may consist of multiple rooms with different requirements for contamination control. Rooms in a clean facility should be maintained at static pressures higher than atmospheric to prevent infiltration by wind. Positive differential pressures should be maintained between the rooms to ensure airflows from the cleanest space to the least clean space. The only exception to using a positive differential pressure is when dealing with specific hazardous materials where governmental agencies require the room to be at a negative pressure. Pressure differentials must be 0.05 in WC higher in an adjacent clean area from the less clean area. This makes airflow critical.

68 Design Considerations and Operation:
Ventilation and makeup air volumes are dictated by the amount required to maintain indoor air quality, replace process exhaust and for building pressurization. This provides assurance that carbon dioxide and oxygen remain in balance, that formaldehyde and other vapors given off by building materials and furniture are diluted, and that air changes occur with sufficient frequency to minimize the chance for high concentration of airborne pollutants within the building. Typically, makeup air must be 15% of all air per air change.

69 Hygiene and Specialized Attire:
Cleanroom environments require people and they also require that those people are clean. Shower or bath- All employees should shower or bath before coming to work. This help to reduce the amount of dead skin cells that are shed when the employee arrives to work. Clean hair is important for reducing flaking from the scalp

70 Hygiene and Specialized Attire:
Clean shaven faces will reduce the potential of contamination from hair that may be exposed in some cases. Smoking before entering a cleanroom will create contamination and should not be done at least thirty minutes before entering. Clothing should be newly cleaned and free of lint. Efforts should be made to reduce contact with animals while wearing clothing that will be worn to work.

71 Hygiene and Specialized Attire:
Hands should be clean and disinfected per the company protocol. Shoes and socks must always be worn. Never wear sandals or open-toed shoes.

72 Preparation to Enter a Cleanroom:
Personal Considerations Hygiene Illness Clean Shaven Smoking Jewelry Make-up Material Considerations Material of Construction Minimal Amounts of Materials

73 Preparation to Enter a Cleanroom:
Disinfection Tools and Supplies Dedicated Tools Hand Washing Material Transfers Flow (separate) Multiple Coverings

74 Gowning (Supplies and Technique):
Hand Disinfectant Safety Glasses Alcohol Beard Cover Gown Surgical Mask Hair Net Sterile Sleeves and Gloves Hood Dedicated Cleanroom Shoes or Boots Goggles Shoe Covers

75 Gowning (Supplies and Technique):
Always follow protocols Disinfect hands prior to gowning Obtain gowning supplies according to protocol Don cleanroom attire from head to toe (literally) except footwear Disinfect hands Aseptically don sterile gloves Obtain footwear Don while crossing over bench Disinfect gloves with alcohol View gowned self in mirror Obtain any other garments before entering (such as sterile sleeves)

76 Cleanroom Behavior: Hygiene- Good basic hygiene is a necessity when working in cleanrooms. Shedding skin cells from unclean workers is the most common contaminant. Bacterial contaminants can also thrive on shed skin cells. Workers who have exudative lesions on their bodies (open wounds with blood or pus) must exclude themselves from cleanroom environments or appropriately and completely cover the sores.

77 Cleanroom Behavior: Coughing or sneezing sometimes cannot be avoided, but it is important to exclude oneself from working in a cleanroom if the cough or sneeze is from and illness or allergy. When coughing or sneezing, always turn away from the area where cleanliness is desired most. Above all, always wear protective masks.

78 Cleanroom Behavior: Facial hair can be shed easily and should
be completely covered when working in cleanrooms. Beard covers can be utilized to accomplish this with beards and mustaches. All masks and beard covers are porous and can still allow contaminants to pass through.

79 Cleanroom Behavior: Motion- All behaviors in cleanroom environments should be performed at a moderately slow pace. Quick movements in the cleanroom or any sterile area stir up the particle counts where they can be easily detected and frequently out-of-specification. Slow, deliberate movements are always best to avoid the turbulence that is created that will stir up particles. Scratching oneself through the cleanroom garments, running, jumping, and even handshakes, are all forbidden practices. It goes without saying that eating, drinking, or smoking is also forbidden.

80 Cleanroom Behavior: Contact- Contacting surfaces is an easy way to
transmit bacteria, viruses, yeast, and molds. If touching any area within a cleanroom by necessity, be sure that you evaluate what you may have “picked-up” and transferred to another area. For example, you may open a door knob or handle and then touch a stopper on a vial that will in theory contain a sterile product. A good practice is to regularly and frequently disinfect your gloves with sterile alcohol between processes. Even a routine disinfection of gloves is appropriate.

81 Cleanroom Behavior: Processes- Obviously, contamination can be
generated by the work we do. For technicians Who are maintaining or repairing equipment, Every mechanical operation performed creates a potential for contamination. Sawing, grinding, fastening screws, hammering, cleaning, drilling, all are sources of contamination. Flames used in cleanrooms, whether being used for sterilization or not, create turbulence which in turn stirs up particles. Even the removal of tools to perform This work will create high particle counts.

82 De-gowning: Reverse the order of gowning and refer to protocol for reuse or disposal of materials.

83 Documentation: Laboratory Notebooks- Any procedure or protocol that is not part of a standard operating procedure should be documented in a laboratory notebook so that it can be referenced at a later time what activities took place and the results that were obtained.

84 Documentation: Data Collection Forms- Provided an SOP is in place that describes the environmental monitoring procedures employed, a data collection form can be used. This form should be referenced in the SOP and designed in an orderly format that allows the manual recording of data and activities that occur during the monitoring procedures

85 Documentation: Electronic Data Collection and Part 11 Compliance- Any electronic data that is obtained as part of the monitoring process must be Part 11 complaint to ensure that data integrity is maintained. Written documentation should be double-checked for accuracy and completeness. All printed materials must be intact and maintained so that they can be archived for future reference. Printed data on thermal paper must be photocopied to preserve the original data that will fade otherwise.

86 Questions? Tour of DSM

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