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6 Operation

38

B-390

 

Operation Manual, Version E

6 .6 

Practicing with the Encapsulator, using alginate solution

After getting comfortable with the bead formation controls, perform test runs with non-sterile algi-
nate solutions. Sodium alginate is the most commonly used polymer, but there are others in use 
with varying properties. We recommend the low viscosity grade alginate. The alginate concentration 
strongly influences the viscosity and this in turn influences the pressure drop in the nozzle. There-
fore, the maximum concentration of the alginate solution is a function of the nozzle diameter (see the 
following table).

Table 6-2: Recommended alignate concentration (based on dry weight) for different nozzle diameters

Nozzle diameter

Concentration of low viscosity grade alginate

Working range

Recommended concentration

80 to 120 

µ

m

0.75 to 1.4 %

1.1 to 1.2 %

120 to 200 

µ

m

1.0 to 1.6 %

1.3 to 1.4 %

200 to 300 

µ

m

1.2 to 1.8 %

1.5 to 1.6 %

300 to 500 

µ

m

1.5 to 2.5 %

1.8 to 2.0 %

NOTE

Under normal storage conditions the alginate powder contains 10 - 12 % water. Therefore we refer to 
the alginate concentration on a dry weight base.

6 .6 .1 

Preparation of 1 .5 % Na-alginate solution

1.  Take a 400 mL beaker and weigh in 3.3 g Na-alginate powder of low viscosity grade.

2.  Add 200 mL of deionized water and mix vigorously with a laboratory mixer for 1 to 2 minutes.

3.  Alginate has the tendency to get lumpy. Remove the alginate lumps from the beaker and the mixer 

blades with a spatula and mix again for 1 to 2 minutes. If lumps remain in the liquid, repeat mixing.

4.  Then let the mixture stand so that the trapped air bubbles will escape from the liquid.

5.  If needed, de-gas the mixture under reduced pressure.

6.  Dissolution of alginate with a magnetic stirrer takes much more time and should be done over-

night.

NOTE

Alginate solutions will support the growth of microorganisms and are stable for about 2 weeks in a 
refrigerator.  An indication of microbial contamination is reduction of the mixture’s viscosity. Alginate 
solutions can be stored for much longer time, even at room temperature, if sterilized or if preserva-
tives are added, like 0.05 % NaN

3

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Содержание Encapsulator B-390

Страница 1: ...Encapsulator B 390 Operation Manual 11593477E en...

Страница 2: ...s the right to make changes to the manual as deemed necessary in the light of experience espe cially in respect to structure illustrations and technical detail This manual is copyright Information fro...

Страница 3: ...ges Table of contents 1 About this manual 5 2 Safety 6 2 1 User qualification 6 2 2 Proper use 6 2 3 Improper use 6 2 4 Safety warnings and safety signals used in this manual 7 2 5 Product safety 9 2...

Страница 4: ...lution 39 6 7 Theoretical background 41 6 7 1 Bead productivity and cell density 43 7 Maintenance and repairs 46 7 1 Customer service 46 7 2 Housing condition 46 7 3 Sealing conditions 46 7 4 Cleaning...

Страница 5: ...required for its safe opera tion and to maintain it in good working order It is addressed to laboratory personnel in particular If the instrument is used in a manner not specified in this manual the p...

Страница 6: ...ecules drugs flavor fragrances pigments extracts cells and micro organisms under open conditions Aseptic working conditions are possible because all parts in contact with the encapsulation mixture are...

Страница 7: ...which if not avoided could result in death or serious injury CAUTION Indicates a hazardous situation which if not avoided may result in minor or moderate injury no NOTICE Indicates possible property...

Страница 8: ...ar laboratory coat Wear protective goggles Wear protective gloves Additional user information Paragraphs starting with NOTE transport helpful information for working with the device software or its su...

Страница 9: ...njuries by use in explosive environments Do not operate the instrument in explosive environments Do not operate the instrument with explosive gas mixtures Before operation check all gas connections fo...

Страница 10: ...syringe pump unit Do not place any laboratory glasses or other utensils on the Encapsulator 2 5 2 Safety measures Always wear personal protective equipment such as protective eye goggles protective cl...

Страница 11: ...erformed with care and on schedule by authorized personnel only Spare parts to be used Use only genuine consumables and genuine spare parts for maintenance to assure good system performance and reliab...

Страница 12: ...livery can only be checked according to the individual delivery note and the listed order numbers NOTE For additional information about the listed products see www buchi com or contact your local deal...

Страница 13: ...ing nozzle rack Pressure bottle 500 mL 11058190 Pressure bottle 1000 mL 11058191 Glass bottles with fittings tubes and air filter working pressure up to 1 5 bar autoclavable Grounding set 11058189 Ope...

Страница 14: ...nded spare parts Table 3 4 Recommended spare parts Product Order no O ring set for single nozzle 11057954 O ring set for concentric nozzle 11057955 Pre filters for nozzle diameter 7 mm 10 pcs 11057957...

Страница 15: ...mL min Maximal allowed air pressure in the system 1 5 bar Reactor gross volume 4 5 liter Reactor working volume 2 liter Parts in contact with medium autoclavable Sterile working conditions limited Ove...

Страница 16: ...ectrostatic Dispersion Unit EDU approximately 5 relative standard deviation of bead size using pure alginate Immediate process control Visual monitoring in the light of a stroboscope lamp High cell vi...

Страница 17: ...ersion E Figure 4 1 Schematic representation of the Encapsulator B 390 a Pressure bottle Bead producing unit Vibration unit Single nozzle Electrode Dispersion control Vibration control LED stroboscope...

Страница 18: ...nozzle size flow rate and physical properties of the encapsulation mixture In general the bead size is twice the nozzle diameter But by varying the jet velocity and the vibration frequency the range...

Страница 19: ...ider the maximum product dimensions and weight The instrument must be set up in such a way that the main switch and the mains plug are easily accessible at all times Obtain the environmental condition...

Страница 20: ...orporated in the control unit Vibration frequency light intensity electrostatic dispersion and heating are controlled on the touch screen Air pressure is regulated with the pressure regulating valve T...

Страница 21: ...sure at the air outlet is 1 5 bar However the working range is from 0 to 1 bar 5 3 Electrical connections Verify that the electrical requirement of the unit stated on the type plate of the control uni...

Страница 22: ...Pre filter with 50 m mesh g O ring 3 68x1 78 h Luer Lock i Screw M3 8 Screw M3 25 Screw M3 6 with attached fixation ring and screws M3 5 You can remove the fixation ring for cleaning Figure 5 3 Parts...

Страница 23: ...h the electrode they pick up the electrostatic charge This electrostatic charge is transferred to the polymerization bath and accumulates if the vessel is not grounded The electrostatic field created...

Страница 24: ...5 Putting into operation 24 B 390 Operation Manual Version E Figure 5 7 Position of the electrode below the carrier plate Figure 5 8 Grounding the polymerization bath...

Страница 25: ...9 Pressure bottle with HEPA filter for sterile pumping of the immobilization mixture with air pressure a Pressure stable flask of 500 mL or 1 000 mL b HEPA air filter c PTFE tube 4 6 d Silicone tube f...

Страница 26: ...ttle with the immobilization mixture 3 Attach the silicone tube of the pressure bottle to the luer lock inlet of the bead producing unit 4 Pass the silicone tube in the liquid regulating flow valve Sq...

Страница 27: ...m and one pressure bottle of 1000 mL The shell liquid is pumped by air pressure using the pressure bottle The main parts of the concentric nozzle unit are refer to figure 5 12 the nozzle pair with she...

Страница 28: ...owing nozzle apertures are standard 0 20 0 30 0 4 0 50 0 60 0 70 and 0 90 mm Figure 5 14 Single parts of the CN bead producing unit a Screw M3 6 b Luer Lock c Pre filter with 50 m mesh d Flat gasket 1...

Страница 29: ...ad producing unit There is no thread The inner nozzle is centered and fixed by the shell nozzle Exit of the shell liquid O ring 12 42 1 78 Figure 5 15 Mounting of the inner nozzle Put carefully the sh...

Страница 30: ...liquid can pass Figure 5 18 Connection of the pressure bottles to the air outlet A T piece feeds both pressure bottles 5 9 Final installation check This check has to be carried out after every install...

Страница 31: ...t breakup control When the Encapsulator is switched on the touch screen runs an initialization program for few seconds Then the screen shows the start menu with four sub parts see figure 6 1 to 6 5 fo...

Страница 32: ...ostatic dispersion parameter The system needs few moments to reach the set value Pressing Esc will return you to the start menu and the set value will be kept Screen 6 3 Electrostatic dispersion unit...

Страница 33: ...a return to the start menu and the set value will be kept Screen 6 5 More options concerning amplitude of vibration and light Intensity of the stroboscope lamp 6 3 Menu structure of the control unit T...

Страница 34: ...lf venting system in the valve The pressure is indicated on the touchscreen see screen 6 1 Note The pressure of air or nitrogen entering the control unit on the rear panel of the Encapsulator should b...

Страница 35: ...800 Hz 5 Open the flow regulating valve by turning the knob counter clock wise until the water flows through the silicone tubing and the bead producing unit to the nozzle where it forms a continuous...

Страница 36: ...lectrostatic tension Clear bead chain with electrostatic tension Lowest frequency Highest frequency Electrostatic Voltage Lowest frequency Highest frequency Nozzle size Air pressure Clear bead chain w...

Страница 37: ...the working range It can happen that after some time the beads no longer enter or actually jump out of the beaker This is due to the fact that electrostatic charges have accumulated in the electrical...

Страница 38: ...0 NOTE Under normal storage conditions the alginate powder contains 10 12 water Therefore we refer to the alginate concentration on a dry weight base 6 6 1 Preparation of 1 5 Na alginate solution 1 Ta...

Страница 39: ...ir pressure still corresponds to the set value Activate the vibration control system and set the vibration frequency at 1100 Hz for the 200 m nozzle and at 800 Hz for the 300 m nozzle 5 Open the liqui...

Страница 40: ...artial occlusion due to dried out polymer mixture 9 Check the beads under a microscope with a micrometer scale eyepiece and record your obser vations of diameter uniformity and shape in table 6 3 10 R...

Страница 41: ...tion mixture It is possible to change opt by 30 and still achieve a good bead formation The diameter of a bead d m can be calculated with the flow rate V m3 s and the frequency of the pulsation accord...

Страница 42: ...the correlation between the vibration frequency and the bead diameter for five different flow rates as calculated by equation 4 Lower flow rates which corresponds to lower pumping rates produce smalle...

Страница 43: ...80 and 120 m nozzles Upper values with application of high voltage NOTE For solutions with a viscosity different from the tested one it can be said that the higher the viscosity the higher the minimal...

Страница 44: ...ion Manual Version E Figure 6 6 Amount of beads with a diameter of 0 3 to 0 6 mm formed from 1 mL of immobilization mixture Figure 6 7 Amount of beads with a diameter of 0 6 to 1 1 mm formed from 1 mL...

Страница 45: ...l Version E Figure 6 8 Amount of cells per bead made from different cell concentrations for bead diameters of 0 3 to 0 6 mm Figure 6 9 Amount of cells per bead made from different cell concentrations...

Страница 46: ...nowledge of possible dangers which might arise when working at the instrument Such training and knowledge can only be provided by BUCHI Addresses of official BUCHI customer service offices are given o...

Страница 47: ...loves 7 4 1 Cleaning the nozzle after each immobilization run It is critical to clean the nozzle immediately after use so that the encapsulation medium alginate etc will not dry and clog the system 1...

Страница 48: ...e in water the appropriate solvent in 1N NaOH or 1N sulfuric acid never use HCl according to the encapsulation mixture for 1 hour at room temperature with periodic agitation Sonication of full stainle...

Страница 49: ...liquid flow rate is too high or too low The Nozzle is not adequately cleaned The amplitude is too low or too high Non homogenous bead size distribution The liquid flow rate is too high The frequency...

Страница 50: ...s e g heating block and carrier plate have cooled down To disassemble the Encapsulator B 390 follow the installation instructions in section 5 in reverse order Remove all liquids and dusty residues be...

Страница 51: ...ter 3 3 This helps to ensure that the components can be separated and recycled correctly You have to follow valid regional and local laws concerning disposal For help please contact your local authori...

Страница 52: ...ause harmful interference in which case the user will be required to correct the interference at his own expense Fran ais Cet appareil a t test et s est av r conforme aux limites pr vues pour les appa...

Страница 53: ...has not been used in the laboratory and is new was not in contact with toxic corrosive biologically active explosive radioactive or other dangerous matters is free of contamination The solvents or res...

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Страница 56: ...w buchi com cn India BUCHI India Private Ltd IN Mumbai 400 055 T 91 22 667 75400 F 91 22 667 18986 india buchi com www buchi in Indonesia PT BUCHI Indonesia ID Tangerang 15321 T 62 21 537 62 16 F 62 2...

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