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Microbiology laboratory building
 Microbiology laboratory building 
Composition microbiology laboratory preparation room, sterilization room, sterile room, and general laboratory incubation chambers of five parts. The common feature of these rooms are floor and wall texture smooth and hard, simple furnishings instruments and equipment to facilitate cleaning. The basic requirements for microbiological laboratories as follows: 
(A) preparation room 
Preparation room for the preparation of medium and sample processing. Indoors there reagent cabinet, storage equipment or materials counter, bench, electric stove, refrigerator and a sewer, power and so on. 
Preparation room is also used for washing utensils. Room should be equipped with heaters, steamer, washing utensils with pots, buckets, etc., there should be a variety of bottle brush, scouring powder, soap, washing powder and so on. 
(B) the sterilization chamber 
Sterilization chamber is mainly used for sterilization sterilization and various appliances medium, room should be equipped with high-pressure steam sterilizer, oven and other sterilization equipment and facilities. 
(C) sterile room 
Sterile room, also known as vaccination room, is a system inoculation, sterile purified laboratory strains and other special operations. Microbial work, bacteria inoculation transplant is a major operation, the characteristics of this operation is to ensure purebred strains to prevent bacteria contamination. General environment in the air, because there are many dust and bacteria, it is easy to cause pollution, work interference on vaccination great. 
1 sterile room settings 
Sterile room should be based on both economic and scientific principles to set. The basic requirements are the following: (a) sterile room should be inside and outside the two, the room is sterile room, outside a buffer chamber. Room volume is not too large, so that air sterilization. Among the smallest in the area of ​​2 × 2.5 = 5m2, outside the area of ​​1 × 2 = 2m2, high to 2.5m below is appropriate, there should be a ceiling. 
(2) should be located within the room sliding door, in order to reduce the volatility of air, the door should be located in a position farthest away from the table; outside with sliding doors at best, to be set up in the room farthest away from the position . 
(3) the separation between the outside of the wall or "sliding door", we should open a small window for the inoculation process necessary internal and external delivery channel of goods, in order to reduce the number of people out of the room, reducing pollution levels. Small window width 60cm, high 40cm, thickness 30cm, both inside and outside to hang on to pull the sash. 
(4) sterile chamber volume is small and tight, after a period of use, the indoor temperature is high, it should be set louvres. Louvres should be located at the inner chamber door on the ceiling (ie furthest away from the table position), the best two-layer structure, the outer shutters, available pumping plate inner sash. After the louvres in the inner chamber may be used to open sterilized prior to air circulation. Constant temperature and humidity conditions can be installed. 
2 sterile room equipment and utensils 
(A) sterile room table, no matter what the material, uses, and require a smooth surface countertops levels. 
(2) the inner chamber and the outer chamber of the installation of a UV lamp (mostly 30W). UV lamps within the chamber to be mounted in a seat are often work above the ground 2m, the outer chamber can be attached to the outer chamber ultraviolet lamp center. 
(3) the outer chamber should have special clothes, shoes, hats, masks, filled with water to the pot Soviet children and towels, hand-held sprayers and 5% carbolic acid solution and so on. 
(4) within the chamber should be alcohol lamp, commonly vaccinated tools, stainless steel knives, scissors, tweezers, 70 percent of the alcohol swab, industrial alcohol, contained in glass, specialty crayons, notebooks, pencils, paper labels, glue, waste baskets and so on.
3 aseptic sterilization chamber 
(1) fumigation: This is completely sterile room sterilization measures. Use sterile room for a long time, pollution is serious, it should be fumigation sterilization. Formaldehyde is available, lactic acid or sulfur fumigation. 
(2) Spray: Before each use, sterile chamber. Spray can contribute to airborne particulate and microbial settlement, to prevent the desktop, dust flying field on the ground, and have a bactericidal effect. Available 5% carbolic acid spray. 
(3) UV irradiation: In the sterile chamber prior to each use. UV has a good bactericidal effect. UV light exposure should normally be open 30 ~ 60min. 
4 sterile room work order 
(A) sterile room sterilization. Before each use of ultraviolet light irradiation 30min turned over, or prior to use 30min, inside and outside the chamber with 5% carbolic acid spray. 
(2) Wash hands with soap after, the equipment needed to move into the outer chamber; outer chamber put sterilized overalls, work cap and work shoes, wearing masks, then phenol soap with 2% coal will hand dip 2 minutes . 
(3) the various items required to move into the room inventory in place, with 5% carbolic acid in the work surface side and the spray operator space station, return the outer chamber, 5 ~ 10min and then work into the inner room. 
(4) before inoculation operation, with 70% alcohol cotton towels; When aseptic action to light 
Slow to minimize fluctuations in air and ground dust. 
(5) work should pay attention to safety. In case of fire tampons, hand grip or wrap off with a damp cloth, do not blow your mouth, so expanding combustion; cases where the bacteria cultures have bacteria spilled or broken container, the application of infiltration 5% carbolic acid rag wrapped after and wipe the table or floor with a rag infiltration 5% carbolic acid with alcohol cotton towels before continuing. 
End (6) work, immediately clean up the table, will not be stored in a sterile room and waste all the items out after the sterile room, sterile room with 5% carbolic acid spray, or open ultraviolet light exposure 30min. 
(Four) incubation chamber 
Setting the culture chamber 1. 
(A) training room should the inner and outer two, the room is the training room, outside the room is a buffer chamber. Room volume should not be large, in order to facilitate air sterilization, the chamber area 3.2 × 4.4 = about 14m2, outer chamber area 3.2 × 1.8 = about 6m2, with about 2.5m high is appropriate, there should be a ceiling. 
(2) the upper part of the wall separating the inner chamber and the outer chamber vents should be located with air filtration devices. (3) In order to meet the need for temperature microorganisms, install constant temperature and humidity. 
(4) inside and outside the chamber should be installed in the central room of ultraviolet light for sterilization. 2 indoor cultivation equipment and utensils 
(A) the chamber is typically equipped aircraft and shake flask cultures machine (shaker). The machine has a conventional rotary flask, reciprocating two kinds. 
(2) the outer chamber should have special clothes, shoes, hats, masks, hand-held sprayers and 5% carbolic acid solution, 70% alcohol cotton ball and so on. 
Culturing chamber 3 sterilization, disinfection 
Sterile room with sterilization, disinfection measures. 
Time to enable small-scale culture incubation room, and in the incubator conducted. 
(Five) ordinary laboratory 
Observation of microorganisms, physiological and biochemical measurements counts and places of work. Interior furnishings and are very different due to the different work focus. Generally equipped with bench, microscopes, cabinets and stools. Bench requires smooth, experimental cabinet should be sufficient to accommodate appliances and medicines and other daily use.

Microbiology Laboratory of major equipment 
 
(A) Clean Bench 
Clean Bench is a local laminar flow device can be formed noble work environment at the local level. It consists of tables, filters, fans, plenum chamber and support other components, the use of filtered air so that the table operating area to purify sterilization purposes. Indoor air through the pre-filter and post-efficiency filter dust vertical or horizontal laminar flow state operating area through the table because there is no air vortex, so any little dirt or dust adhering to the bacteria can be ruled out, easy to elsewhere proliferation and metastasis. Accordingly, the operating area can remain sterile. 
Compared with the sterile room and inoculation box, use clean bench has good working conditions, easy to operate, reliable and sterile effect, no harm to human body disinfectants, small footprint and can move and so on. If placed in a sterile indoor use, sterile better results. The disadvantage is expensive, pre-filter and high efficiency filters also need regular cleaning and replacement. 
(B) high-pressure steam sterilization pot 
High-pressure steam sterilizer is a closed, can withstand a certain pressure double metal pot. Within a water pot or mezzanine, when the water in the pot to boil because of the steam can not escape, so the pressure gradually increased the pot boiling point of water and temperature will be increased, so as to achieve the purpose of high-temperature sterilization. Generally carried out at a pressure 0.11MPa, 121 ℃ sterilized 20 ~ 30min, including all microorganisms, including spores can be killed. If sterilization items larger, steam penetrate difficulty, an appropriate increase in steam pressure sterilization or extend the time. 
Autoclave horizontal, vertical, and other types of portable, in microbiology laboratories, the most commonly used is portable and vertical high-pressure steam sterilization pot. And pressure autoclaves, compared the advantages and autoclave 
Points are required for a short time sterilization, save fuel, thorough sterilization and so on. The disadvantage is that expensive, smaller sterilization capacity. 
(C) incubator 
Incubator is to develop microorganisms special equipment. Heating type incubator is heated by a heating wire fixed volume and temperature control device synthetic training device, the size of the different specifications. Microorganism commonly used laboratory incubator working volume with 450 × 450 × 350mm3 or 650 × 500 × 500mm3, applicable to all types of room temperature to 60 ℃ between the microbial culture. Now, with the development of science standards, sophistication and price incubator devices are very different. There are a variety of reasonable structure, complete functions of the incubator, such as incubator, constant temperature and humidity incubator, low temperature incubator, microbiology and more carbon dioxide incubator incubator and so on. Some with computer control, you can choose multiple timelines transformation temperature, thus overcoming the effects of ambient temperature can reach temperatures all year round training requirements. 
Microbial incubator is a multi-use heating, cooling and vibration in one of the microbial liquid fermentation device. Studio arbitrarily selected temperature within the range of 15 ~ 50 ℃, the temperature controller is selected by the automatic control, keep working indoor temperature. SCR also features speed control system, oscillation machine speed can be arbitrarily regulated within 1 ~ 220rpm range. 
(Four) oven 
Oven is a device for removing wet materials and utensils inside and outside water or other volatile solution. Many types, there are boxes, drum, suite-style, rotary and so on. Microbiology Laboratory Multi-box oven, the size of the different specifications. Working indoors with movable fence panels, easy to place the dried goods. Heating heating type electric oven is also composed of silk and temperature control device can adjust the temperature from room temperature to 300 ℃ arbitrary choice. Some oven thermometer sensitive element using a conductive, transistors and relays with automatic control system to overcome the shortcomings of thermal expansion of the metal tube to control. 
In addition, a vacuum oven (with a vacuum pump and a pressure gauge), can be operated at atmospheric or reduced pressure. 
(Five) Shakers 
Also known flask shaker machine, which is to develop small-scale test equipment aerobic microorganisms or as expanding culture of seeds, commonly used in reciprocating rotary shaker has two. Reciprocating a reciprocating shaker frequency is generally 80 to 140 times / min, the stroke is generally 5 ~ 14cm, such as the frequency is too fast, the stroke is too large or too much the amount of the bottle of liquid mounted in the liquid shaking bottle wrapped splash gauze or cotton stuffed, resulting in bacterial contamination, especially when this happens it easier to start. 
Eccentricity of the rotary shaker in general between 3 ~ 6cm, the rotation rate of 60 ~ 300rpm. In shaker flasks (typically flask) of oxygen in the fermentation broth is determined by the required air through the mouthpiece gauze dressing (typically 8 layers), or through the cotton plug, so the oxygen transfer and bottle the size, geometry, bottle, or gauze tampon thickness and density. In general, the absorption coefficient of oxygen flask sample loading volume depends on the characteristics and shaking the flask. 
Reciprocating shaker shaker drive is to use the principle of a crank for reciprocating movement, the body is made ​​of iron or wood framed rectangle, a layer to three pallets, are prepared circular tray placed on the flask, a protruding hole triangular rubber vibration reduction for fixed flasks and bottles, the transmission mechanism generally use two pulleys deceleration, change speed pulley can change reciprocating frequency. Opening holes have different eccentric cam to adjust the eccentricity. The frequency and size of the reciprocating shaker eccentricity significantly influence the absorption of oxygen. 
Using a rotary shaker rotating eccentric shaft swings the tray, the tray with one or two layers, can be stainless steel, aluminum or wood board manufacturing. In three eccentric shaft with adjustable bolt down the tray remains level. This shaker structural complexity and high cost. The advantage is a better oxygen transfer, power consumption is small, the medium is not splashed on the bottle gauze.
(Seven) inoculation box 
Inoculation box is divided into two boxes inoculated solid and liquid inoculation. Inoculation is a solid box made ​​of wood and glass or plexiglass welded together by a closed casket. Divided into double and single operation box. Box vary, generally the box is about 143cm, width 86cm, total height 154cm, stand 76cm. The upper left and right sides of each box can be fitted with two leaves of glass sliding doors open and close, easy to strain out. The lower part of the window, respectively, with two round hole about 13cm in diameter, the center distance between the two holes of 52cm (shoulder width), the hole is equipped with elastic cuff, to prevent your hands when operating in the box, outside air into the tank the cause pollution. They sit opposite operation, his hands through the cuff extends into the box. Both sides of the box is also fitted with the best glass top boxes for wood or glass. Top box is equipped with ultraviolet germicidal lamps and fluorescent lighting each one. Cabinet install wood or glass can, but be careful sealed. 
Inoculated liquid tank is designed for transposing liquid spawn and design. Bacteria than solid elongated box, unilateral two operations. Features tracks and ultraviolet light, both ends of the box to open a high 25cm, width 10cm rectangular outlet, convenient bacteria and out the hole with a small door. Import and export Xiachu set a vapor source, import and export of seal inoculation with steam to prevent the bacteria entering the box. The back of the box has moved to take over the liquid bacteria can enter the hole. 
Inoculation box sterilized with ultraviolet irradiation 30min. If there is no ultraviolet light, can be used formaldehyde and potassium permanganate (formaldehyde 10 ~ 14mL / m3 + potassium permanganate 5 ~ 7g / m3 space) fumigation than 30min. When used, the first vaccination required items and tools into the box, and then fumigated and UV sterilization agents, and then aseptically inoculated. 
Simple inoculation box structure, low cost, easy sterilization, easy to operate, and people operating outside the box, can work at higher temperatures. The disadvantage is that the medium access time and labor, are required prior to each vaccination sterilization. 
(Eight) refrigerator 
Microbiological laboratory refrigerators are mainly two types: ordinary refrigerators and cryogenic refrigerator. Ordinary refrigerator generally have two cabinets, namely fresh Tibetan cabinets and freezers, temperatures were 4 ℃ and -20 ℃; cryogenic refrigerator temperature control at -40 ~ -80 ℃. They can be used for the Deposit of Microorganisms. Fresh Tibetan cabinet 
 
Commonly used to save slant bacteria, preserved time in three months. More than 3 months, the slope will become dry, and therefore needs to transfer species. If you want to save the species for a long time, you need to go through post-processing, storage in an ordinary refrigerator freezer or cryogenic refrigerator, they preserved a long time, generally more than a year. 
 
 
Microbiology Laboratory Management 
 
A laboratory management system 
1. Laboratory instruments should be developed with the management, use the system, drug management, use the system, glassware management, the use of the system, and in accordance with requirements of safety systems and environmental conditions, this room staff should be strictly controlled, carefully executed. 
2. Into the lab must wear overalls, non-laboratory personnel shall not enter the laboratory, strict implementation of safety procedures. 
3. Laboratory items neatly, agents regularly checked and have clear labeling, instrumentation regular inspection, maintenance, overhaul, is strictly prohibited in the refrigerator for storing and processing of private food. 
4. Various equipment should be established for claiming consumption records, precious instrument has to use records, broken missing report should be completed; medicines, equipment, bacteria are not allowed to borrow without the approval and transfer, and more shall come privately. 5. Smoking is prohibited in the laboratory, dining parlor, noise, may not bring personal items in the laboratory, water and electricity before leaving the laboratory carefully check for toxic, hazardous, flammable, pollution, corrosive substances and waste materials should be relevant Requests the Executive. 
6. Strict implementation of the person in charge of the system, report the problem immediately. 
Second, the instrument is equipped, management and use of the system 
1. Food microbiology laboratories should have the following equipment: incubator, pressure cooker, ordinary refrigerator, cold refrigerator, anaerobic training equipment, microscopes, centrifuges, clean units, oscillators, general balance, one-thousandth of balance, oven, freeze-dried equipment, homogenizer, thermostatic water bath, colony counter, incubator, potential pH meter, high-speed centrifuges. 
2 laboratory instrument used, the container shall comply with the standard requirements to ensure accurate and reliable, all measuring instruments shall be certified by the measurement department before using. 
3. Laboratory instruments placed reasonable, there is someone keeping valuable instrument to establish the instrument archives, and with methods of operation, maintenance, repair, and use of manual registration of this, do regular maintenance, maintenance and inspection, precision instruments are not free to move, if damaged when in need of repair, not secretly dismantled, should write the report, notify managers, by the manager agreed to fill repair application, send equipment maintenance department. 
4. Various instruments (refrigerator, temperature except me), immediately after use to cut off the power knob recovery homing, to be carefully examined before leaving. 
5. All instruments and equipment without equipment managers agree not borrow, after the registration of the use by content were registered. 
6. Equipment should be kept clean, it should normally instrument shroud. 
7. When using the instrument should be carried out strictly in accordance with the rules, in violation of the rules of the instrument due to mismanagement resulting in equipment damage, the parties concerned should be held accountable. 
Third, drug management, use system 
1. The room inspection tasks in accordance with the development of various pharmaceutical agents procurement plan, include full name, unit, quantity, purity, packaging specifications, date of manufacture, and establishing accounts, after someone managed to reclaim every six months to make the consumption table, and the remaining inventory drugs. 
2. Drug agents display tidy, orderly place, away from light, moisture, air dry, bottle label intact, locked storage of highly toxic drugs, flammable, volatile, corrosive species separate storage. 
3. Consuming drugs reagents, please fill out the required lead single, and room by using the person in charge to sign any person not entitled to privately 
Lend or feed drug agents, the unit Branch, Room occasionally signed by the department head of interlibrary loan from other units when needed. 4. Weigh Drug agents should be operating specifications, after using cover, if necessary, seal or black paper parcel, do not use expired or deteriorated drugs. 
Fourth, glassware management, use the system 
1. According to the requirements of test items, glassware declaration procurement plan, detailing the specifications, origin, quantity, requirements, hard neutral glass instrument should be verified by qualified metering. 
2. Large containers establish accounts, inventory once a year, usually low-value consumable inventory at any time after completing the registration utensils damage loss. 
3. After glassware dirt should be removed before use, and use cleaning fluid or 2% diluted hydrochloric acid solution soak for 24 h, rinse with water reserve. 
4. After cleaning utensils to use at any time after the contamination should be strictly autoclaving, not indiscriminate disposal of litter. 
Fifth, security system 
1. Into the laboratory work clothes, hats, shoes must be worn neatly. 
2. During high pressure, drying, disinfection and other work, the staff are not allowed from the scene, carefully observe the temperature, time, distilling volatile, flammable liquid when you are not allowed to directly heat the water bath should be set, such as produced during the test in the cabinet should avoid poison gas operations. 
3. When the draw is strictly prohibited drugs and bacteria directly with the mouth, according to the aseptic operation, such as bacteria occur, the pathogen spill containers, shall immediately be thoroughly disinfected with an effective disinfectant, before leaving the scene safe handling. 4. After work, wash hands with soap and water, if necessary, can Bromogeramine, peracetic acid foam hand, then rinse with water, clothes should be cleaned regularly to keep clean and, if necessary, autoclaved. 
5. Experiment is completed, the immediate cleaning up the site and experimental equipment for contamination infected items, disinfection and sterilization process. 6. Daily work, especially around holidays, carefully check the water, electricity and equipment being used, shut the doors and windows before leaving. 
Six, environmental conditions 
1. The laboratory should always be kept clean, cleaning should be carried out every day to work on finishing, desk cabinets and other surfaces should be 
Wipe with a disinfectant every day, to keep clean, to prevent contamination. 2. The laboratory should orderly, shall not be stored outside the laboratory and personal belongings, equipment and other laboratory supplies to put reasonable, and have a fixed position. 3. Keep in laboratory health not throw confetti and other debris, test the waste to be used in a fixed box down the barrel, and timely manner. 
4. The laboratory should have excellent light conditions and lighting. 
5. Laboratory work surface should be level and no leakage, walls and floors should be smooth and easy to clean. 6. Laboratory layout to be reasonable, generally between laboratories should prepare and sterile room, sterile room should have good ventilation conditions, such as installation of air conditioning equipment and filtration equipment, sterile indoor air testing should be basically sterile. 7. Do not use the laboratories for conference rooms and other recreational activities and learning spaces.