RTS-1C — User manual
Contents
1. 3 3 A 40 W External power Supply cc cseeeeeeeeeeeeeeeeeaeees Input AC 100 240V 50 60Hz Output DC 12V Optional accessories Catalogue number USB 2 0 Hub 10 ports BS 010158 BK Biosan is committed to a continuous programme of improvement and re serves the right to alter design and specifications of the equipment without addition al notice i 0 25 OD A600 nm equivalent isi 0 20 OD A600 nm equivalent or Accurate within 10 18 9 1 D2 9 3 9 4 Maintenance If the unit requires maintenance disconnect the unit from the mains and contact Biosan or your local Biosan representative All maintenance and repair operations must be performed only by qualified and specially trained personnel Standard ethanol 75 or other cleaning agents recommended for cleaning of laboratory equipment can be used for cleaning and decontamination of the unit Clean the rotor of the device from liquid droplets and possible contamination after finishing fermentation 10 10 1 10 2 10 3 10 4 10 5 10 6 10 7 10 8 10 9 20 Device calibration for maintenance Note Only use this calibration if the measurement system does not work correctly Turn on the device Insert a 50 ml centrifuge tube filled with 10m of H20 into the device Set the Volume parameter to 10ml Using control panel or software unit control panel set RPM to 2000 Press Run Stop button Press and hold Select button until CC2. command will appear and blink on the display Press A button and 0 00 number will appear and blink on the display Wait 15 seconds and press W button Device is now calibrated and will make one verification measurement 11 Warranty and Claims 11 1 The Manufacturer guarantees the compliance of the unit with the require ments of Specifications provided the Customer follows the operation stor age and transportation instructions 11 2 The warranted service life of the unit from the date of its delivery to the Customer is 24 months Contact your local distributor to check availability of extended warranty 11 3 If any manufacturing defects are discovered by the Customer an unsatisfac tory equipment claim shall be compiled certified and sent to the local distrib utor address Please visit www biosan lv Technical support section to obtain the claim form 11 4 The following information will be required in the event that warranty or post warranty service comes necessary Complete the table below and retain for your records Model Real Time Cell Growth Logger with cooling RTS 1C Date of sale Serial number 21 12 Declaration of Conformity 22 23 Biosan SIA Ratsupites 7 build 2 Riga LV 1067 Latvia Phone 371 67860693 371 67426137 Fax 371 67428101 http www biosan lv Version 1 01 January 2015
3. of prokaryotes and their classification I Psychrophiles 1 2 facultative Il Mesophiles Ill thermophiles 3 thermotolerant 4 facultative 5 obligate 6 extremophile Thick line mark represents optimal growth temperature 7 1 1 For psychrophiles the device must be installed in a cold room or refrigerated chamber Despite the active cooling device the sample is heated through the open upper part of falcon tube Moreover it also should be considered that the actual temperature of the reactor will always differ from the actual temperature of the sample because of its rotation at low temperatures below 10 C and will be higher 7 1 2 For mesophilic microorganisms the device can be situated at room tempera ture 7 1 3 For thermophilic microorganisms please observe Fig 4 where comparison of thermoblock temperature real temperature in the tube depending on rota tion intensity can be observed Real T C inside the tube Fig 4 Comparison of thermoblock temperature real temperature in the tube depending on rotation intensity The end point data was collected 1 30h after start of the heating process from room temperature 14 7 2 Cell growth depending on rotation intensity It is known that aeration affects the growth and growth rate of aerobic microorganisms The reverse spin frequency affects the rate of oxygen up take in the bioreactor Results obtained indicate that the maximum rate of cell
4. the subsequent proce dures Take a 50 ml falcon tube Add 10 0 1ml distilled water Close the cap of the falcon thoroughly Insert the falcon into the socket fig 1 4 Set the volume parameter of the distilled water on the display fig 2 11 Press Run Stop key fig 2 9 the device will start the OD measurement cycle by accelerating to 2000 rpm The measurement cycle should complete after 15 20 seconds and OD value should appear on the display If OD value equals 0 0 05 OD then the device corresponds to factory pre calibration settings and is suitable for use 10 2 Operation Recommendations during operation ol 92 5 3 5 4 O 5 6 5 9 5 10 Remove the falcon tube from the tube socket before connecting or discon necting the external power supply during operation It is recommended to start operation approximately 15 minutes after switch ing on the device the time necessary for stabilization in the working mode Connect external power supply unit to electric circuit fig 1 1 Turn on the unit by pressing the power switch on the rear panel fig 1 3 Insert the tube into the socket fig 1 4 Press the Select key fig 2 1 to choose a parameter that you want to change the active parameter is blinking Use A and keys fig 2 2 to set the necessary value if the key is pressed for more than 2 seconds the parameter should change quicker It is possible t
5. CHD Medical Biological Research amp Technologies RTS 1C Real Time Cell Growth Logger for version V 1AW Operating Manual Certificate Contents pl Oy UO sek I Se 10 11 12 Safety Precautions General Information Getting Started Calibration Operation Approved methods for microorganism cultivation Recommendations for creating personal settings for cultivation of microor ganisms Points that should be considered 7 1 7 2 1 3 17 4 17 4 7 5 Temperature distribution specifics osychrophiles mesophiles thermophiles Cell growth depending on rotation intensity Aeration and types of recommended tubes Cell growth depending on measurement frequency aerobes anaerobes Particle size and calibration coefficients 600nm 850nm Linear range of OD measurement as a function of medium volume Specifications Maintenance Device calibration for maintenance Warranty and Claims Declaration of Conformity 1 Safety Precautions The following symbol means A Caution Make sure you have fully read and understood the present Manual before using the equipment Please pay special at tention to sections marked by this symbol GENERAL SAFETY Use only as specified in the operating manual provided The unit should not be used if dropped or damaged After transportation or storage keep the unit under room temperature for 2 hrs before connecting it to the electric circuit Use onl
6. division is detected at a frequency of 1 Reverse Spin per second 1 s at a speed of 2000 rpm The increase of pause between reverse spins re duces cell growth rate reaching 50 of the maximum value when RS fre quency is 30 s see Fig 5 and Fig 6 OD over tme OO GO GA eS RS Oe Ores 7 howd E s 0 5U 70 Hours U srowth rate Fig 5 Influence of Frequency of Reverse Spinning on the Growth kinetics AOD A 850nm At vs Time of fermentation hrs 7 2 1 Legend of experiment Fig 5 Real Time Cell Growth Logger RTS 1C was used with 850 nm LED volume of LB media in 50 ml Falcon tube is approx imately 15 ml Reverse Spin Frequency 1 2 4 8 16 30 s measurement frequency MF is approximately 10 mint reactor rotation speed 2000 rpm temperature 37 C diameter of filter pores for aeration 0 25 um 1 3 16 Growth rate Fig 6 Influence of Frequency of Reverse Spinning on the Growth kinetics AOD A 850nm At vs Time of fermentation hrs Aeration and types of recommended tubes For aerobic microorganisms we recommend using tubes that are supplied by us TPP TubeSpin Bioreactor 50ml but for aerotolerant anaerobes like Lactobacillus acidophilus it is necessary to use test tubes with sealed lids non membrane For obtaining optimal results growing aerotolerant anaer obes it is required to seal the screw cap of TPP TubeSpin Bioreactor 50ml by tape User can also use standard ce
7. e in testing vessel 10 min but not less Measurement Frequency MF Thermophilic aerobe Thermophilus sp 2000 rpm 1s RSF 70 C Real tube temperature 5 C lower 15 ml 10 min MF Evaporation rate at 70 C 5 ml 24 h please adjust Volume parameter ac cordingly for measurement system to work correctly Aerotolerant anaerobe L acidophilus O rpm 0s RSF 37 C 30 ml 10 min MF We recommend using Profiling Cycling preset No mixing OD meas for homogeneous distribution of cell suspension in the bioreactor tube prior to measurement Currently studies are being conducted to better understand the advantages of reverse spin RS technology for growing different types of microorgan isms such as yeast and lactic acid bacteria It is possible for the end user to contact the manufacturer for advising or suggesting a required microorganism or strain for cultivation to be tested Please contact the R amp D department of Biosan at these e mail adress sci ence biosan v igor biosan v Igor Bankovsky consulting biotechnologist on application questions 699 7 Recommendations for creating personal settings for cultivation of microorganisms Points that should be considered 7 1 Temperature distribution specifics osychrophiles mesophiles thermophiles The optimal growth temperatures of microorganisms are divided three prin cipal groups see Fig 3 Fig 3 Temperature boarders and optimal growth zones
8. eaaeeeeeeeaaaeeeeseaaaeeeeeeaaaeeeeeeeaaes 1 pce Operating Manual Certificate cccccccccseseecceceeeeeeeeeeeeeeeeseaeeeeseeees 1 copy Optional accessories USB hub with 10 POMS sceececetsccicesnncrscncdexsanrestvencetesteentnasteetebeet lt secet on request 4 z RTS 1C 11 f a E en o e biosan Fig 1 Rear panel Fig 2 Control panel 3 3 Set up Place the unit on even horizontal working surface Connect the external power supply unit to the socket fig 1 1 on the rear side of the unit Switch on the computer if it was turned off Install the software following the software installation procedure described in software installation manual Bioreactor vessel features Falcon type tubes TPP TubeSpin Bioreactor 50 type tubes are required to use the kid with openings Working volume 10 30 ml Conical form 5 openings A B C D E of different size above the gas permeable sterile PTFE filter of the screw cap Openings can be sealed and by this exchange adjusted to need Steril gas exchange is guaranteed by the 0 22 um filter membrane Even with a high cell density the supply of oxygen through the openings is sufficient Tube fits in a standard 50 ml centrifuge rotor 4 Calibration The device is pre calibrated at the factory for operation with 50ml falcon at temperature range from 4 C to 70 C and saves calibration data when being switched off To verify the conformity of calibration follow
9. ntrifuge tubes of 50 ml Falcon type taking into account that the tube will be as transparent as TPP TubeSpin Bioreactor tube 7 4 Particle size and calibration coefficients 6 00nm 850nm Calibration of the instrument is designed for specific microorganism size of 0 4 0 8 x 1 3 um and a cell volume of about 0 6 0 7 um In case of exceed ing the allowable size the measurement system will not work correctly Optical density OD A 850 nm to OD A 600 nm conversion rate coefficient is equal to 2 2 Example of calculation to convert 3 5 OD A 850 nm to OD A 600 nm simply multiply the result by 2 2 resulting in 7 7 OD A 600 nm 7 5 Linear range of OD measurement as a function of medium volume Fig 7 Volume influence on RTS 1 OD measurement linearity Fig 7 Influence of Broth medium volume on linear range of OD measurement AOD A 850nm At vs Time of fermentation hrs As can be seen from Fig 7 the device registers optical density of the sample in linear range until 15 OD A 600 nm at a volume of 10 ml 8 OD A 600 nm at 20 ml and 6 OD A 600 nm at 30 ml 8 Specification The unit is designed for operation at ambient temperature from 4 C to 40 C in a non condensing atmosphere and maximum relative humidity 80 for temperatures up to 31 C decreasing linearly to 50 relative humidity at 40 C 8 1 Measurement specifications LAGIAS OUNG Gica anette assert ers oe ease ec ear IR LED Wavelength A MIE ren
10. o E E E E 850 15 Measurement range OD at 10 20 iil VOM E asni E O 10 00 at 20 30 MI VONMIMNG xc sssiac hess seesn as ipeeiuscete T Seesiec ieeewsieteens 0 8 00 Measurement precision OD ccccccceecceceeecceceeeceeseeeeeseeeeeeseeeeeeseneeeeas 0 3 Real time measurement MIN MEASULEMEN ccecceeseeeeeeeeeeeeeeeaeeees 1 60 TIME setting resolution MIN c ccc cccsecccceeeeceeeeceeeeeeeeeeseeeeseueessaeessaeeeseeeesaes 1 8 2 Temperature specifications SUING range Clera E tc ee eet eee 4 0 Bottom control range point C cccceeccessseeeesseeeeeseeeeeseees 15 below ambient Lop control Tange DOU CO secede eerie cies e eee itactuatseeisranntiatat 0 Seting resol tion O x co rece ears snce mentee torts ce a a tees conteanees 0 1 SLAIN oo cscscceclet ss hesstaa cocaine Mazes saciaincpastendecte iia Vases tedtcihaanteaseatesees 0 1 8 3 General specifications Recommended max number of units connected to the software 12 SAMPIOVOIUME IMM is cclaeilt cer a cee a a aaah 10 30 Speed FANGS TOM aicitendeed nsgeilins haven endian 50 2000 Speed setting reSOlUtlON TOM eses i a 1 BE Fe PEET E PEENE EEE E EEE EE ME a E E SE EAE EE REE E E TNE LCD Overall dimensions W x D x H MM ccccseeeeesseeeeeeeeeeeeeees 130x212x200 Woon e KO ienaa chin dane et send See een ee neces ce re aetece ee acandeemieee 1 5 Input current power CONSUMPTION eeeee eee 12 V DC
11. o set time between optical density measurements fig 2 3 spinning speed fig 2 4 temperature fig 2 5 time between Reverse Spins fig 2 8 operating volume fig 2 11 Actual values of the temperature and speed are displayed on the display fig 2 6 and fig 2 7 Note After turning on the unit starts heating and continues to maintain the temperature regardless of other operations Press the Run Stop key fig 2 9 to start and stop operation Press the Run Stop key fig 2 9 to stop the operation Caution Operation stop will not stop the heating process To stop heating process set temperature has to be decreased manu ally until off indication appears fig 2 5 After finishing the operation switch OFF the unit with the Power switch fig 1 8 Disconnect external power supply unit from electric circuit fig 1 1 6 Approved methods for microorganism cultivation We are completely aware that the world of microorganisms is infinite and each strain requires its own growth optimization New methods for cultivation of microorganisms using RTS 1C technology will with no doubt increase and we will inform our customers about it Here are the 3 approved methods that were developed by Biosan application lab 6 1 6 2 6 3 6 4 6 5 12 Facultative anaerobe Escherichia Coli 2000 rpm vessel spinning speed 1 s Reverse Spin Frequency RSF 37 C socket temperature 15 ml sample volum
12. onsibility to carry out appropriate decontamination if hazardous material is spilt on or penetrates into the equipment 2 General Information Real Time Cell Growth Logger is a device which can be used as an incuba tor with optical density measurement in real time Temperature control allows using RTS 1C as an incubator for example to cultivate cells Due to innovative mixing technology reverse spinning of the sample around its own axis it is possible to make OD measurement without probe interference Developed software for recording displaying and data analysis allows working in real time Real Time Cell Growth Logger is equipped with a cooling unit that allows cooling the samples to 4 C and temperature profiling through sofware The Real Time Cell Growth Logger is applicable in e Molecular biology e Cell biology e Biotechnology e Biochemistry e Chemistry e Microbiology Features e Innovative mixing due to reverse spinning of the tube with the sample around its Own axis e Due to innovative mixing technology it is possible to measure optical density and light scattering of the sample in real time without probe interference maintaining process sterility e Changing the parameters such as temperature revolution per minute and period of spinning in one direction together with possibility of creating exper iment algorithms including temperature profiling mixing intensity profiling optical density control etc allow
13. s both performance of difficult sequence of algorithms of fermentation process and achieving consistent and repeatable results Software possibilities Remote tracking and control of fermentation process Real time registration of cell growth kinetics or particle suspension aggrega tion disaggregation processes User graphs including 3D graphs Pause Saving and loading result data PDF and Excel spreadsheet reports Simultaneous connection of up to 10 units that allows on one hand to research the influence of different chemical and physical factors on fermen tation process and on the other hand to research the interdependance of those factors in matrix experiments 3 1 3 2 Getting started Unpacking Remove packing materials carefully and retain them for future shipment and storage of the unit Examine the unit carefully for any damage incurred during transit The warranty does not cover in transit damage Complete set The unit set includes Standard set RTS 1C Real Time Cell Growth Logger with COoling ccccccseeeeee 1 pce B a ete Sate a sere ete ee Soar meinen E E O E AE spe ener E 1 pce Bioreactor vessels TPP TubeSpin Bioreactor 50ml c 0000ee 20 pcs Dala AO arase e AE 1 pce USB disk drive with software installation files and installation MANUAI cccecccseececeececeeeeceeeeeseececeeeessueeeseeeseeeeseaees 1 pce External power Supply cccccccceeeceeseeeeec
14. y cleaning and decontamination methods recommended by the manufacturer Do not make modifications in design of the unit ELECTRICAL SAFETY Do not plug the unit into the main socket without grounding and do not use extension lead without grounding Connect only to a power supply with voltage corresponding to that on the serial number label Use only the external power supply unit provided with this product Ensure that the external power supply unit and switch are easily accessible during use Disconnect the unit from the electric circuit before moving Turn off the unit by switching off the power switch and disconnecting the external power supply unit from the power socket If liquid penetrates into the unit disconnect it from the external power supply unit and have it checked by a repair and maintenance technician Do not operate the unit in premises where condensation can form Operating conditions of the unit are defined in the Specifications section DURING OPERATION e Do not operate the unit in environments with aggressive or explosive chemi cal mixtures Please contact manufacturer for possible operation of the unit in specific atmospheres e Do not operate the unit if it is faulty or has been installed incorrectly e Do not use outside laboratory rooms e Do not check the temperature by touch Use a thermometer e Always clean and decontaminate the rotor after operation BIOLOGICAL SAFETY e It is the users resp
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