APPLICATION NOTE Orbital shaker benchmarks: best practices for use and maintenance

Authors: Mary Kay Bates, Senior Global Cell Culture Specialist and Sara Livingston, Global Product Manager, Thermo Fisher Scientific

Key words: Orbital shaker, HEPA filter, cleaning, orbit diameter

INTRODUCTION Most life science labs have an orbital shaker—often more than one— because these versatile workhorses are used in many applications including , biochemistry, molecular biology, microbiology, environmental and food sciences, clinical diagnostics, and eukaryotic Figure 1. Orbital shakers are available in a wide variety of sizes and formats. cell culture. Usage extends from basic research all the way up to maintenance schedule. In all cases, for before discarding any packing bioproduction. Shakers come in the manufacturer’s recommendations materials; some small parts can be a wide range of sizes and formats for each model take precedence and accidentally discarded. (Figure 1), including open air, heated, should be followed. or refrigerated. Choosing among Recognize that orbital shakers, these many options can be confusing, INSTALLATION regardless of size, can be heavier than and your application may require a Upon delivery of your new shaker, you may think. Take care when lifting specific shaker style1. inspect the shipping carton and the shaker, ensure you have at least ask the carrier to specify and sign one other person to help you, and that Regardless of the models used, this for any damage on your delivery the shelf or bench designated to hold application note contains valuable receipt, to ensure that any damage the shaker will be strong enough. information which applies to all to the components can be properly orbital shakers. We offer helpful documented and addressed. When tips to properly install your shaker, unpacking any equipment, guidelines for best operation, proper carefully match parts to a packing list usage and cleaning, and a general and ensure everything is accounted

1 Also confirm that the shaker will not the shaker to a power source until platform. Consider a shaker which wobble or tip. Orbital shakers can it is completely set up and ready offers platforms which are easily vibrate if not properly supported or for use. With larger shakers we installed and removable to allow for while experiencing an uneven load. recommended setting up each unit easy cleaning (Figure 2). In general, a Vibration can have additional adverse on an independent circuit to provide shaker platform will have integrated effects on the mixing motion created, stable power to the unit and avoid points to aid in alignment of the impacting cell growth and the power fluctuations that may affect the platform over the drive mechanism. end product. electronics of the shaker system. Install all screws before tightening them, to ensure proper seating of the LOCATION For best operation, use a level and platform. Once installed, never lift Because shakers vary so much in adjust turn screws on the unit feet to the shaker by the platform, but only size, the required clearance around ensure the shaker is level (including by the bottom and sides. Use only the particular model will also vary. front to back as well as side to side). the manufacturer provided screws, But all shakers should have some Tighten the locking nuts to secure tools, and clamps. Establish a clear, space around them to allow for the position. organized and well labeled area, proper ventilation. In general, this ideally a clear plastic divided box, to space should be 1 inch (2.5 cm) Depending on your shaker make store these parts near the shaker. for smaller units, up to 4 inches and model, you may have to install (10 cm) for the largest models. Also, the shaker platform next. Before Before installing flask clamps, check because the shaker is working to progressing too far, however, know whether a flexible spring must first establish a set temperature, it is that some shakers require the flask be installed. If your clamps require important to position the shaker clamps to be manually secured onto a spring, it must be used, because out of areas of direct sunlight or the platform before installing the without it the flask will not be secured drafts. The shaker’s temperature control can also be influenced by nearby equipment, especially any that generate heat, such as ultra-low freezers or ultracentrifuges.

If you have a refrigerated shaker, it may need access to a drain for evacuating condensed liquid. Some shakers have a sealed chamber with a drain for convenience when cleaning with liquids. This drain may be hidden behind an access panel. If you plan to use this drain, consider its location when positioning the unit.

INITIAL SETUP The shaker may have a switch next to the power cord receptacle to toggle between 115 or 230 volts. Set the Figure 2. Removable platforms are recommended for all orbital shakers for greater flexibility switch appropriately for the voltage and to facilitate regular cleaning and disinfection. A universal platform will accept a wide used in your region. Do not connect variety of clamps.

2 and there is a great risk of the flask breaking, spilling, or accidentally releasing from the clamp. After attaching the springs, install the clamps onto the platform using the manufacturer provided instructions and parts. Different shakers and flask clamps require 1–5 screws, and some require nuts and/or washers, so be sure to use all of the necessary components for your model. Using fewer than the recommended number Figure 4. Clamps are available that are designed to fit Thermo Scientific™ Universal of screws and nuts will increase the Racks. The clamps can rotate to allow incubation at a desired angle. Different rack models fit risk of the flask clamp dislodging from a variety of commonly used culture tubes. The racks easily pop into and out of the clamps. the platform, causing damage to your flask and samples. alarms—to more closely match your holder and take the entire rack and most common settings. cultures with you to and from the If you are connecting a data shaker, eliminating the need to transfer monitoring and remote alarm module, Ideally, the shaker will have a tubes to a new rack for transport. now is a good time to set that up. This display that shows all parameters These are so easy to use, save time, is also time to become familiar with simultaneously (Figure 3) and allows and eliminate the waste and risk of the different alarms indicated by the precise individual adjustments. using laboratory tape to secure test shaker and how to correct them. If Refer to the user manual for proper tube racks to the shaker platform. your shaker allows, you can change management of settings, and become the alarm setpoints and triggers— familiar with any specific tips for We recommend establishing a such as high and low temperature routine parameter entry. standard procedure to check the platform and flask clamp screws TIPS FOR BEST OPERATION monthly and tighten if necessary, to AND CELL PRODUCTION prevent failure and breakage. When placing a flask into a spring clamp, use your free hand to pull the Load flasks and tubes into clamps and clamp open enough to easily insert racks carefully and securely, making the flask. This will increase the service sure all are firmly seated before life of the clamp and spring, and startup. When using multiple vessels, prevent accidental breakage of the ensure that they are evenly distributed flask. Reverse the process to remove on the platform for best operation and the flask, again using one hand to pull to reduce the chances of vibration, the spring and clamp open before uneven wear on mechanics, and pulling the flask out. uneven shaking.

For shaking culture tubes, consider Always begin with the shaker set to using a specialized “0” RPM (revolutions per minute). Figure 3. A shaker should offer a clear holder (Figure 4) which will secure the Increase the shaking speed gradually display which allows viewing of all parameters simultaneously and allows rack and tubes at a specified angle. to the desired speed, monitoring to individual, precise control. You can remove the rack from the ensure you get a “swirl” action as

3 the speed increases. While this will least once each month. Normal indoor take longer than simply switching the air contains thousands of circulating 2 d1 shaker on and allowing the speed to r2 = r1 × microorganisms which can take up increase as fast as possible, it is the d2 residence in your shaker, putting your best approach because increasing the cultures at risk. speed too quickly can create a “slosh” Here, d1 = the orbital diameter for rather than the gentle swirling that is the first shaker, d2 = the diameter for At a minimum, take a good look at ideal for best mixing and cell viability. the new shaker, r1 = the RPM for the the shaker parts and remove any

This is most critical for animal and first shaker and r2 = the RPM for the spilled growth media or dirt using a insect cells which do not have a cell new shaker3. microfiber cloth moistened in water wall; however, this procedure will aid with mild dish soap. Rinse with clear in protecting all cell types and liquid Different speed, temperature, flask water. Using a spray , wet all solutions. Often, you can use a slower size and shape, liquid volume, cell parts and areas with 70% ethanol speed once you have achieved a types, and growth media components and allow to air dry. Do not spray any good swirling motion around the flask all have effects on cell growth. In most areas with exposed electronics. perimeter. In addition, by personally cases any negatives you experience monitoring the shaker until it reaches with cell growth will require some Remember that spills can seep under the set speed, you will be there to empirical testing and comparison of the platform. Orbital shakers which stop the shaker if any problem occurs different variables. To best maintain offer easily removable platforms during acceleration, and to correct the conditions inside an incubated ease cleaning of this often neglected the situation. or refrigerated shaker, try to limit the area. Larger models may offer a frequency and duration of shaker lid drain to facilitate the process. For If the above procedure does not give or door openings. large incubated/refrigerated floor you the cell viability or productivity models, HEPA filtration is a feature you need, experiment with a new Never operate the shaker at maximum that automatically purifies the air, flask with a different aspect ratio (the speed without a load, or overload saving you time and protecting your ratio between the height of the flask the systems weight limits. The stated cultures from contamination. Filters compared to its diameter), or one with maximum weight for a shaker will be are generally good for 12 months, can baffles. Also, test different culture the combined weight of the platform, be autoclaved with other biological lab volumes; this is especially important clamps, vessels, and samples. waste and are easy and inexpensive for scale up because the volume to to replace. flask physics play a significant role in CLEANING AND DISINFECTION cell health. Best mixing and aeration Clean up any spills immediately using Some laboratory shakers offer are achieved when the liquid is a lint free cloth dampened with a ultraviolet (UV) light as a mode 10–25% of the flask volume2. noncorrosive cleaner or 70% ethanol, of decontamination. We do not or as instructed by the manufacturer. recommend using this option because Shakers are available with various This is important to keep liquids from we do not believe it is effective orbital diameters; however, with the entering the shaker mechanism or enough for standard use in an orbital appropriate speed, flask size and electronics cabinet. In addition to shaker environment. UV light is volume, any size orbit will provide attending to sudden spills, regular easily blocked by even dust and dirt, sufficient aeration for proper cell cleaning is also important, for best rendering it ineffective. Areas not growth. Remember that shakers shaker operation and to protect your directly reached by the light—such as with different orbit sizes will affect cultures from contamination. We on the shadowed side of clamps—are the speed required, according to the recommend cleaning and manually not disinfected. In practice, UV light following equation: disinfecting your laboratory shaker at efficacy decreases as the bulb ages

4 and in many cases those using UV plastic to protect the cover surface. light are unaware that it is no longer 3. If your shaker has a touch screen adequate. In contrast, HEPA filtration display, use a dry microfiber is continuously working and becomes cloth. Use liquid cleaners only as more efficient over time as the filter approved by the manufacturer. collects particles. The HEPA filter will 4. If your shaker has an air filter capture all particle sizes, although it is (Figure 5), remove and inspect it least efficient—99.97%—at 0.3 µm. for dust and dirt. It may be cleaned with mild soap in water, and dried REGULAR PREVENTIVE between two lint free towels. MAINTENANCE 5. Inspect under the shaker unit and Like most laboratory equipment, clean up any dust, dirt or debris. your orbital shaker will perform at its 6. Ensure that the shaker platform best when it is properly maintained. and all clamps are properly seated Refer to the user manual for your and secured. model to ensure you are following the manufacturer’s recommendations. Quarterly Figure 5. A typical air filter and cover for a Here are some general guidelines: 1. If your shaker offers refrigeration, refrigerated shaker. cleaning the coil and Monthly air filter is recommended every 1. Unplug the unit and clean inside and 3 months. Generally these can be filter. This may be required more out using a mild dish soap and wipe cleaned using mild dish soap in frequently depending on the dry. You may follow with 70% ethanol water and thoroughly dried with a usage of the unit and surrounding as a disinfectant. Do not allow lint free cloth. ambient conditions. detergent or disinfectant to make 2. Verify the proper operation of the contact with any electrical systems. circulating fan. More specific recommendations Remove and clean under the shaker 3. If your shaker has a belt based can be found in the user manual platform if possible. If you require a mechanism, visually inspect the drive for your model. Contact a qualified stronger disinfectant, we recommend belt for signs of wear. Refer to your service professional for questions a quaternary ammonium based user manual for specific instructions or service of shaker belts, probes, product such as Lysol™ No Rinse on maintaining and replacing belts. fans or electronics. In general, sanitizer, Conflikt, or Fermacidal D. shakers available today should not Quaternary ammonium is safe for Yearly require any oiling. Thermo Fisher stainless steel, plastic and glass, and 1. Test and document the accuracy of Scientific factory trained service will not leave harmful fumes which the temperature calibration, speed, professionals are available in could affect your cultures. time and alarms. Also check the most areas and can be found at

2. For cleaning an acrylic shaker cover calibration of CO2 (if applicable). Use www.unitylabservices.com. you can use any plastic cleaner; be secondary probes and sensors that aware that some glass cleaners can have been themselves calibrated cause clouding of acrylic, so make in the last year to eliminate any sure to use one recommended for variability. Position the secondary plastic. Use only a soft cotton or probe in the center of the shaker microfiber cloth; never use a scouring chamber and allow the unit time to compound or pad. You may use a equilibrate to the setting. polish specifically recommended for 2. If applicable, change the HEPA

5 CONCLUSIONS Table 1. Typical maintenance list and schedule for an orbital shaker. Refer to the user manual for each model for specific recommendations. Orbital shakers are found in most and despite their frequent Maintenance schedule summary for orbital shakers usage are often overlooked in terms Action Monthly Quarterly Yearly of standard maintenance. With proper Manually clean and disinfect the shaker. installation and care, they should • provide years of trouble free service. Inspect the condenser air filter. Clean as needed. An ideal shaker is easy to use and • program and is easily assembled Inspect underneath the shaker and remove any dust or debris. • and disassembled to allow user- Check that the shaker platform and clamps are friendly maintenance. properly seated and secured. • Clean refrigeration condenser coil and air filter. REFERENCES • 1. O’Bryan J, Goodman T (2001) Choosing the right orbital Verify proper fan operation. shaker for your application. Thermo Fisher Scientific • ANSHKSELECTOR 0611. If applicable, inspect the drive belt for signs of wear. 2. Hensler H-J and Schedel M (1991) Suitability of the • shaking flask for oxygen supply to microbiological Test accuracy of temperature, speed, time and alarms. cultures. Bioprocess Engineering 7, 123–131. • 3. Bates MK, Phillips DS, O’Bryan J (2016) Shaker agitation rate and orbit affect growth of cultured bacteria. Thermo If applicable, change to a new HEPA filter. Fisher Scientific ANSHORBGRT 0816. •

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