Operation Manual
Encapsulator
B-395 Pro
11593484
en
Table of contents
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
2.5.1 General hazards. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .9
2.5.2 Safety measures . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10
2.5.3 Built-in safety elements and measures . . . . . . . . . . . . . . . . . . . . . . . . 10
2.6 General safety rules . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
2.7 Disclaimer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11
3 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.1 Scope of application and delivery . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.1.1 Standard instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12
3.1.2 Standard accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13
3.1.3 Optional accessories . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
3.1.4 Recommended spare parts . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
3.2 Technical data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
3.3 Materials used. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15
4 Description of function . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
4.1 Functional principle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16
4.2 Connections at the Encapsulator B-395 Pro. . . . . . . . . . . . . . . . . . . . . . 18
5 Putting into operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.1 Installation site. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19
5.2 Installing the Encapsulator B-395 Pro . . . . . . . . . . . . . . . . . . . . . . . . . 20
5.3 Electrical connections . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21
5.4 Assembling of the reaction vessel . . . . . . . . . . . . . . . . . . . . . . . . . . . 22
5.4.1 Cover plate . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23
5.4.2 Base plate. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27
5.4.3 Bead collecting flask . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29
5.5 Pumping systems . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
5.5.1 Syringe pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
5.5.2 Pressure bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31
Read this manual carefully before installing and running your system and note the safety precautions
in chapter 2 in particular. Store the manual in the immediate vicinity of the instrument, so that it can be
consulted at any time.
No technical modifications may be made to the instrument without the prior written agreement of
BUCHI. Unauthorized modifications may affect the system safety, the EU conformity or result in accidents.
This manual is copyright. Information from it may not be reproduced, distributed, or used for competitive purposes, nor made available to third parties. The manufacture of any component with the aid of
this manual without prior written agreement is also prohibited.
The English manual is the original language version and serves as basis for all translations
into other languages.
3 B-395 Pro Operation Manual, Version C
5.5.3 Installation of the pressure bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . 32
5.6 Option: Concentric nozzle system . . . . . . . . . . . . . . . . . . . . . . . . . . . 33
5.6.1 Mounting of CN nozzles . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35
5.7 All parts of the Encapsulator B-395 Pro . . . . . . . . . . . . . . . . . . . . . . . . 36
5.8 Final installation check. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36
6 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
6.1 Starting up the instrument. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
6.2 Main screens . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37
6.3 Menu structure of the control unit . . . . . . . . . . . . . . . . . . . . . . . . . . . 38
6.4 Menu functions of the upper touch screen . . . . . . . . . . . . . . . . . . . . . . 39
6.5 Menu functions of the lower touch screen . . . . . . . . . . . . . . . . . . . . . . . 41
6.5.1 Menu for syringe pump calibration. . . . . . . . . . . . . . . . . . . . . . . . . . . 42
6.5.2 Selecting a calibrated syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43
6.6 Manual air pressure control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
6.7 Handling the syringe pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
6.7.1 Calibrating the syringe pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
6.7.2 Selecting a pre-calibrated syringe . . . . . . . . . . . . . . . . . . . . . . . . . . . 45
6.8 Practicing with the Encapsulator, using water . . . . . . . . . . . . . . . . . . . . . 46
6.8.1 Using the syringe pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46
6.8.2 Using the pressure bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49
6.9 Practicing with the Encapsulator, using non-sterile alginate solution . . . . . . . . . . 52
6.9.1 Preparation of 1.5 % Na-alginate solution . . . . . . . . . . . . . . . . . . . . . . . 52
6.9.2 Working with the syringe pump . . . . . . . . . . . . . . . . . . . . . . . . . . . . 53
6.9.3 Working with the pressure bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . 56
6.10 Practicing with the Encapsulator, working with the complete reaction vessel . . . . . 58
6.11 Heat sterilization of the reaction vessel . . . . . . . . . . . . . . . . . . . . . . . . 60
6.12 Sterilization of the pressure bottle . . . . . . . . . . . . . . . . . . . . . . . . . . . 60
6.13 Encapsulation procedure for immobilization of micro-organisms in Ca-alginate beads. 61
6.14 Encapsulation protocol for alginate-PLL-alginate membranes . . . . . . . . . . . . . 63
6.15 Theoretical background . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 66
6.15.1 Bead productivity and cell density . . . . . . . . . . . . . . . . . . . . . . . . . . . 68
7 Maintenance and repairs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
7.1 Customer service . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
7.2 Housing condition . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
7.3 Sealing conditions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 71
7.4 Cleaning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 72
7.4.1 Cleaning the nozzle after each immobilization run . . . . . . . . . . . . . . . . . . . 72
7.4.2 Cleaning a clogged nozzle. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 73
7.4.3 Cleaning the reaction vessel and the other vessels . . . . . . . . . . . . . . . . . . 73
8 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
8.1 Malfunctions and their remedy. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 74
9 Shutdown, storage, transport and disposal . . . . . . . . . . . . . . . . . . . . . . . . 75
9.1 Storage and transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 75
9.2 Disposal. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 76
10 Declarations and requirements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
10.1 FCC requirements (for USA and Canada) . . . . . . . . . . . . . . . . . . . . . . . 77
10.2 Health and Safety Clearance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 78
10.3 Declaration of conformity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 79
Table of contents
4 B-395 Pro Operation Manual, Version C
1 About this manual
This manual describes the Encapsulator B-395 Pro. It provides all information required for its safe
operation 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 protection provided by the
instrument may be impaired.
Abbreviations
EPDM Ethylene Propylene Dimonomer
FEP Fluorelastomer
PTFE Polytetrafluoroethylene
1 About this manual
5 B-395 Pro Operation Manual, Version C
2 Safety
This chapter points out the safety concept of the instrument and contains general rules of behavior
and warnings from direct and indirect hazards concerning the use of the product.
For the user's safety all safety instructions and the safety messages in the individual chapters shall
strictly be observed and followed. Therefore, the manual must always be available to all persons
performing the tasks described herein.
2.1 User qualification
The instrument may only be used by laboratory personnel and other persons, who on account of
training and professional experience know the dangers, which can develop when operating the instrument.
Untrained personnel or persons, who are currently being trained, require careful supervision by a qualified person. The present Operation Manual serves as a basis for training.
2 Safety
2.2 Proper use
The Encapsulator B-395 Pro has been designed and built as laboratory instrument.
The Encapsulator B-395 Pro is a semi-automated instrument used for the polymer encapsulation of
chemical substances, bio-molecules, drugs, flavor & fragrances, pigments, extracts, cells and microorganisms under sterile and non-sterile conditions. The bead formation is based on the fact that a
controlled, laminar liquid jet is broken into equally sized beads, if vibrated at an optimal frequency.
The Encapsulator B-395 Pro provides just such controlled conditions to generate beads between 0.15
to 2 mm. The instrument is ideally suited to encapsulate particles < 50 µm.
If the instrument is used with potentially toxic or hazardous substances, it has to be installed inside a
closed fume hood or glove box. In such case, the complete processing and system handling has to
be performed within the ventilated box to avoid toxication and other hazardous situations to the user
and the environment.
2.3 Improper use
Applications not mentioned in section 2.2 are considered to be improper. Applications which do not
comply with the technical data (see section 3 of this manual) are also considered to be improper.
The operator bears the sole risk for any damages or hazards caused by improper use!
The following uses are expressly forbidden:
• Installation or use of the instrument in rooms, which require ex-protected instruments.
6 B-395 Pro Operation Manual, Version C
2.4 Safety warnings and safety signals used in this manual
DANGER, WARNING, CAUTION and NOTICE are standardized signal words for identifying risk levels,
related to personal injury and property damage. All signal words, which are related to personal injury
are accompanied by the general safety sign.
For your safety it is important to read and fully understand the below table with the different signal
words and their definitions!
Sign Signal word Definition Risk level
DANGER
Indicates a hazardous situation which, if not avoided, will result in
death or serious injury.
2 Safety
★★★★
WARNING
CAUTION
NOTICE
no
Supplementary safety information symbols may be placed in a rectangular panel on the left to the
signal word and the supplementary text (see below example).
Space for
supplementary
safety
information
symbols.
Table of supplementary safety information symbols
The below reference list incorporates all safety information symbols used in this manual and their
meaning.
Indicates a hazardous situation which, if not avoided, could result
in death or serious injury.
Indicates a hazardous situation which, if not avoided, may result
in minor or moderate injury.
Indicates possible property damage, but no
practices related to personal injury.
!
SIGNAL WORD
Supplementary text, describing the kind and level of hazard/risk seriousness.
• List of measures to avoid the herein described, hazard or hazardous situation.
• ...
• ...
(property damage only)
★★★☆
★★☆☆
★☆☆☆
Symbol Meaning
General warning
Electrical hazard
7 B-395 Pro Operation Manual, Version C
Explosive gases, explosive environment
Harmful to live-forms
Device damage
Pressurized gas/air
2 Safety
Wear 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
supplementaries. NOTEs are not related to any kind of hazard or damage (see example below).
NOTE
Useful tips for the easy operation of the instrument/software.
8 B-395 Pro Operation Manual, Version C
2.5 Product safety
Safety warnings in this manual (as described in section 2.4) serve to make the user alert and to avoid
hazardous situations emanating from residual dangers by giving appropriate counter measures.
However, risks to users, property and the environment can arise when the instrument is damaged,
used carelessly or improperly.
2.5.1 General hazards
The following safety messages show hazards of general kind which may occur when handling the
instrument. The user shall observe all listed counter measures in order to achieve and maintain the
lowest possible level of hazard.
Additional warning messages can be found whenever actions and situations described in this manual
are related to situational hazards.
2 Safety
!
Warning
Death or serious injuries 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 for correct installation.
• Directly withdraw released gases and gaseous substances by sufficient ventilation.
!
Warning
Pressure increasing in the inlet-system due to clogged nozzles.
Bursting of the inlet system.
Death or serious poisoning by contact or incorporation of harmful substances at use.
• Clean nozzle immediately after use, see section 7.4.
!
Warning
Death or serious injuries by contact with high voltage.
• Only open the housing of the product when machine is switched off and unplugged.
Risk of instrument short-circuits and damage by liquids.
• Do not spill liquids over the instrument or parts of it.
• Wipe off any liquids instantly.
• Ensure a safe positioning of the sample vessel.
• Do not move the instrument when it is loaded with liquid.
• Keep external vibrations away from the instrument.
9 B-395 Pro Operation Manual, Version C
Notice
2 Safety
Risk of instrument damage by wrong mains supply.
• External mains supply must meet the voltage given on the type plate.
• Check for sufficient grounding.
Risk of damaging labratory glasses or utensils by moving 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 clothing, and
gloves when working with the instrument.
2.5.3 Built-in safety elements and measures
High voltage and electrostatic charges
• Safety current limitation.
• Internal grounding to arrest electrostatic charges.
Notice
Notice
Air/Gas
• Over pressure safety valve (opens at 1.5 bar)
10 B-395 Pro Operation Manual, Version C
2.6 General safety rules
Responsibility of the operator
The head of laboratory is responsible for training his personnel.
The operator shall inform the manufacturer without delay of any safety-related incidents which might
occur during operation of the instrument. Legal regulations, such as local, state and federal laws
applying to the instrument must be strictly followed.
Duty of maintenance and care
The operator is responsible for the proper condition of instrument at use and that maintenance,
service and repair jobs are performed 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 reliability. Any modifications to the spare parts used are only allowed with the prior
written permission of the manufacturer.
Modifications
Modifications to the instrument are only permitted after prior consultation with and with the written
approval of the manufacturer. Modifications and upgrades shall only be carried out by an authorized
BUCHI technical engineer. The manufacturer will decline any claim resulting from unauthorized modifications.
2 Safety
2.7 Disclaimer
Use and marketing of any material produced with the Encapsulator are in the sole responsibility of the
operator.
11 B-395 Pro Operation Manual, Version C
3 Technical data
This chapter introduces the reader to the instrument and its specifications. It contains the scope of
delivery, technical data, requirements and performance data.
3.1 Scope of application and delivery
The Encapsulator B-395 Pro is available
• for sterile working conditions in a closed reaction vessel
• with one integrated syringe pump.
The scope of delivery 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 dealer.
3 Technical data
3.1.1 Standard instrument
Table 3-1: Standard instrument
Product Order no.
Encapsulator B-395 Pro
50 – 60 Hz, 100 – 240 V
Encapsulator B-395 Pro
50 – 60 Hz, 100 – 240 V
with GMP documentation
Complete Encapsulator B-395 Pro system for sterile procedures with integrated syringe pump, magnetic stirrer and
closed reaction vessel.
11058220
11058230
12 B-395 Pro Operation Manual, Version C
3.1.2 Standard accessories
3 Technical data
Table 3-2: Standard accessories
Product Order no.
Reaction vessel 11057890
Reaction vessel
11057879
with GMP documentation
Completely autoclavable reactor made of
glass and stainless steel for the sterile
production and collection of microcapsules, 2 litre working volume
Set of 8 single nozzles 11057918
Set of 8 single nozzles with high precision
opening of 0.08, 0.12, 0.15, 0.20, 0.30,
0.45, 0.75 and 1.00 mm, made of stainless steel 316L including 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
Operation Manual English 11593484
13 B-395 Pro Operation Manual, Version C
3.1.3 Optional accessories
3.1.4 Recommended spare parts
3 Technical data
Table 3-3: Optional accessories
Product Order no.
Concentric nozzle set 11058051
Set of 7 external nozzles with high precision opening of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7
and 0.9 mm made of stainless steel, incl.
1000 mL pressure bottle
Table 3-4: Recommended spare parts
Product Order no.
O-ring set for single nozzle 11057954
O-ring set for concentric nozzle 11057955
O-ring set for reaction vessel 11057970
Pre-filters for nozzle,
diameter 7 mm (10 pcs.)
Drain filters for reaction vessel,
diameter 35 mm (10 pcs.)
11057957
11057958
14 B-395 Pro Operation Manual, Version C
3.2 Technical data
Table 3-5: Technical data Encapsulator B-395 Pro
Power consumption max. 150 W
Connection voltage 100–240 VAC
Mains supply voltage fluctuations up to ±10% of the nominal voltage
Frequency 50/60 Hz
Fuse 3.15 A
Dimensions (W×H×D) 32×38×48 cm
Weight 11 kg
Nozzle diameter of single (= core) nozzles 0.08, 0.12, 0.15, 0.20, 0.30, 0.45, 0.75 and 1.00 mm
Nozzle diameter of shell nozzles 0.20, 0.30, 0.40, 0.50, 0.60, 0.70 and 0.90 mm
Droplet size range 0.15 to 2.00 mm
Vibration frequency 40 to 6,000 Hz
Electrode tension 250 to 2,500 V
Syringe pump rate 0.01 to 50 mL/min
3 Technical data
Pump rate by air pressure 0.5 to 200 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 full
Overvoltage category II
Pollution degree 2
Environmental conditions:
Temperature 5–40 °C for indoor use only
Altitude up to 2000 m
Max. relative humidity (curve parameter) Maximum relative humidity 80 % up to 31 °C, then
Table 3-6: Material and Approvals
Material in contact with sample stainless steel, silicone, glass, FEP, PTFE
Approvals CE, CSA
3.3 Materials used
decreasing linearly to 50 % relative humidity at 40 °C
Table 3-7: Materials used
Component Material description
Reactor Stainless steel, 3.3 boroscillate glass, FEP, PTFE
sealings: silicone, EPDM
Nozzles Stainless steel, sealings: EPDM
Pressure bottle Stainless steel, 3.3 boroscillate glass, FEP, PTFE
sealings: silicone, EPDM
15 B-395 Pro Operation Manual, Version C
4 Description of function
This chapter explains the basic working principle of the Encapsulator B-395 Pro. It also shows how
the instrument is structured and provides a general functional description of its assembly.
4.1 Functional principle
The instrument provides the following key functions:
Sterile working conditions in a closed reaction vessel
– Sterile containment in an autoclavable reaction vessel.
Reproducible bead size from one production to the next
– Adjustable parameters (nozzle size, liquid flow rate and vibration freequency) determine bead
size.
4 Description of function
Reproducible bead formation
– In the range of 0.15 mm to 2.0 mm.
High bead size uniformity
– Due to the integrated Electrostatic 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 viability
– Bead formation technique is at low shear stress and under physiological conditions, thus
resulting in high cell survival.
Batch size
– When using syringes the batch size is of 2 mL to 60 mL and the dead volume is approximately
0.5 mL.
dead volume is approximately 2 mL.
Set of 8 single nozzles
– The 8 nozzle sizes of 0.08, 0.12, 0.15, 0.20, 0.30, 0.45, 0.75 and 1.0 mm cover the bead size
range of approximately 0.15 mm to 2.0 mm.
Delivery of the polymere mixture
– By the integrated syringe pump or by air pressure with flow rates from 70 mL/h (0.08 mm
nozzle) to 2’500 mL/h (1.0 mm nozzle).
When using air pressure for pumping the batch size is of 5 mL to 1’000 mL and the
High bead production
– Up to 6000 beads are produced per second depending on encapsulation conditions and
polymer mixture.
16 B-395 Pro Operation Manual, Version C
4 Description of function
14
13
2
1
11
5
6
12
4
7
8
3
10
9
15
17
16
18 19
Figure 4-1: Schematic representation of the Encapsulator B-395 Pro
a Syringe pump
Syringe
Pressure bottle
Air Pressure control
Pulsation chamber
Vibration system
Nozzle
Electrode
Reaction vessel
Bypass cup
17 B-395 Pro Operation Manual, Version C
Liquid filter
Air filter
Electrostatic charge generator
Frequency generator
Stroboscope lamp
Filtration grid
Bead collecting flask
Magnetic stirrer
Waste port
4 Description of function
The main parts of the Encapsulator B-395 Pro are the control unit, with the syringe pump, the electrical and pneumatic systems, and the reaction vessel. All parts of the instrument which are in direct
contact with the beads can be sterilized by autoclaving.
The product to be encapsulated (cells, microorganisms, or other biologicals and chemicals) is mixed
with an encapsulating polymer (typically alginate) and the mixture put into a syringe or a pressure bottle , see figure 4-1. The polymer-product mixture is forced into the pulsation chamber
by either a syringe pump a or by air pressure . The liquid then passes through a precisely drilled
nozzle and separates into equal size droplets on exiting the nozzle. These droplets pass through an
electrical field between the nozzle and the electrode resulting in a surface charge. Electrostatic
repulsion forces disperse the beads as they drop to the hardening solution.
Bead size
The bead size is controlled by several parameters including the vibration frequency, amplitude, nozzle
size, flow rate, and physical properties of the polymer-product mixture. In general, the bead diameter
of Ca-alginate beads is twice the nozzle diameter. But, by varying the jet velocity and the vibration
frequency, the range can be adjusted by about ±15 %.
Optimal parameters for bead formation are indicated by visualization of real-time bead formation in the
light of a stroboscope lamp . When optimal parameters are reached, a standing chain of droplets is
clearly visible. Once established, the optimal parameters can be preset for subsequent bead production runs with the same encapsulating polymer-product mixture. Poorly formed beads, which occur at
the beginning and end of production runs, are intercepted by the bypass cup .
Depending on several variables, 50 to 5000 beads are generated per second and collected in a
hardening solution within the reaction vessel . Solutions in the reaction vessel are continuously
mixed by a magnetic stir bar to prevent bead clumping. In addition, the reaction vessel and/or
solution must be electrically grounded. At the conclusion of the production run, the hardening solution
is drained off (waste port ), while the beads are retained by a filtration grid . Washing solutions,
or other reaction solutions, are added aseptically through a sterile filter . The beads can be further
processed into microcapsules, or transferred to the bead collecting flask .
4.2 Connections at the Encapsulator B-395 Pro
Front connections (See figure 5-2)
• Main switch
• Air out
• Voltage
• Ground
Rear connections (See figure 5-1)
• Electric supply
• Air inlet
• Magnetic stirrer
• Vibration
• Optional plug
18 B-395 Pro Operation Manual, Version C
5 Putting into operation
This chapter describes how the instrument has to be installed. It also gives instructions for the initial
startup.
NOTE
Inspect the instrument for damages during unpacking. If necessary, prepare a status report immediately to inform the postal company, railway company or transportation company. Keep the original
packaging for future transportation.
5.1 Installation site
Put the instrument on a stable, horizontal surface. Consider 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 conditions as described in section 3.2 “Technical data”.
5 Putting into operation
!
Warning
Death or serious injuries by use in explosive environments.
• Do not operate the instrument in explosive environments.
!
Warning
Death or serious poisoning by contact or incorporation of harmful substances.
• Wear safety goggles.
• Wear safety gloves.
• Wear a laboratory coat.
• Clean the instrument and all accessories thoroughly to remove possibly dangerous
substances.
• Do not clean dusty parts with compressed air.
• Store the instrument and its accessories at a dry place.
19 B-395 Pro Operation Manual, Version C
5.2 Installing the Encapsulator B-395 Pro
Place the instrument on the lab bench with convenient access to an AC electrical outlet and to
compressed air. Place the instrument in a way that disconnection of the electric supply plug is
possible at all times.
Installation of the magnetic stirrer, vibration unit and grounding wire
Connect the magnetic stirrer, the vibration unit and the grounding wire as shown in figure 5-1 and 5-2.
1
3
2
5 Putting into operation
a Air inlet (blue tube 2.6×4.0 mm)
Electric supply socket with inte-
grated fuse
4
5
Optional socket
Socket for magnetic stirrer
Socket for vibration unit
Figure 5-1: Rear view of the control unit
All controlling systems for bead production are incorporated in the control unit. Vibration frequency,
pump speed, light intensity, electrostatic dispersion and magnetic stirrer speed are controlled on the
two touch screens. Air pressure is regulated with the pressure regulating valve. The integrated stroboscope lamp allows real time jet breakup control. The vibration unit is attached to the control unit on the
rear panel by a wire. The reaction vessel is attached to the reactor holder with two screws.
a Syringe pump
Reactor holder
Vibration unit
Upper touch screen (vibration
frequency & electrode)
Lower touch screen (syringe
pump, magnetic stirrer control &
pressure indication)
Pressure regulating valve
Stroboscope lamp
Mains switch
Magnetic stirrer
Air outlet
EDU (Voltage outlet)
Plug for grounding wire
LIquid flow regulating valve
13
11
10
1
3
7
2
4
5
6
12
9
Figure 5-2: Front view of the control unit
8
20 B-395 Pro Operation Manual, Version C
Installation of the air line
A 3 m air tube (2.6×4.0 mm) is included with each Encapsulator to connect it to external compressed
air or nitrogen.
1. Stick the air tube into the air inlet plug.
2. Attach the other side of the air tube to the external gas supply.
3. Deliver gas to the Encapsulator at 1.5 to 2 bar (23 to 30 psi) when running the instrument.
NOTE
The integrated pneumatic system (valve and fittings) will tolerate up to 7 bar (100 psi) at the inlet.
An over pressure safety valve, which opens at 1.5 bar, is incorporated after the pressure regulating
valve, so that the maximum air pressure at the air outlet is 1.5 bar. However the working range is 0 to
1 bar.
5.3 Electrical connections
5 Putting into operation
Verify that the electrical requirement of the unit, stated on the type plate of the control unit, corresponds to voltage of your local electrical network. Connect the power plug of the Encapsulator to the
mains supply.
Caution
!
Risk of instrument damage by wrong mains supply.
• External mains supply must meet the voltage given on the type plate.
• Check for sufficient grounding.
• Additional electrical safety measures such as residual current brakers may be necessary to
meet local laws and regulations! External connections and extension lines must be provided
with an earthed conductor lead (3-pole couplings, cord or plug equipment). All used power
cords shall be equipped with molded plugs only to avoid risks due to unobservant defective
wiring.
21 B-395 Pro Operation Manual, Version C
5.4 Assembling of the reaction vessel
1
5
3
9
4
2
8
10
11
16
18
15
12
13
14
17
20
19
6
7
The Reaction vessel forms a closed, autoclavable unit in which the beads are formed under sterile
conditions and can be further processed if neede
5 Putting into operation
a Liquid filter
Connection to electrode
Screw M4×10
Flange
Glass cylinder
O-Ring (6×2) for gap control
Harvesting valve
Plastic clamp
Silicon tube (6×9) of drain
line
Air filter
Bead producing unit
Bypass knob
Syringe
Cover plate
Bypass cup
Support bar
Filter of drain line
Flat silicone fitting
Base plate
t Foot
Figure 5-3: General view of the reaction vessel
The reaction vessel‘s main parts are:
1. Stainless steel cover plate with electrode, bypass, liquid inlet, and air exchange filter
2. Bead producing unit
3. Nozzle
4. Glass cylinder
5. Stainless steel base plate with bead and drain valve
6. Bead collecting flask
22 B-395 Pro Operation Manual, Version C
5.4.1 Cover plate
The cover plate is delivered with all pieces in place. Before use, wash the cover plate carefully. After
each run, disassemble the bead producing unit and the nozzle. Wash with water or appropriate detergent or solvent (according to the nature of the immobilization mixture used), rinse with water and let
dry. Be careful not to damage the PTFE membrane while handling the bead producing unit.
The other parts should be disassembled only as needed. Wash with detergent, rinse with water and
let dry.
When reassembling, check the integrity of the flat fittings and o-ring seals – replace if needed.
1
2
5 Putting into operation
a Front notch for alignment
Groove for fitting
Nozzle
5
6
Liquid inlet
Electrode
Screw M4×12
Bead bypass
3
4
Figure 5-4: Bottom view of mounted cover plate
1
7
a O-Ring 117.1×3.53
Air inlet
O-Ring 29.87×1.78
Liquid inlet
2
3
4
Figure 5-5: Bottom view (left) and top view (right) of cover plate
23 B-395 Pro Operation Manual, Version C
5.4.1.1 Bead producing unit and nozzles
2
3
5 Putting into operation
a Magnet Holder *
41
5 6
O-Ring (14×1.78)
Pulsation chamber
Pre-filter with 50 µm mesh
O-Ring (3.68×1.78)
Luer Lock
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.
Do not remove either the
magnet or the glass fiber
reinforced PTFE membranes!
7
Figure 5-6: Parts of the bead production unit
98
A high quality nozzle is crucial for homogenous bead production. The holes of the Encapsulator
nozzles are precisely drilled using the newest technology. Every Encapsulator B-395 Pro is delivered
with a set of 8 nozzles; nozzle aperture sizes are 80, 120, 150, 200, 300, 450, 750 µm and 1.0 mm.
They are made completely of stainless steel.
Figure 5-7: Set of 8 nozzles on the nozzle rack
The nozzle rack contains 8 nozzles (80, 120, 150, 200, 300, 450, 750 µm, and 1.0 mm). The size of
the O-ring is 4.47×1.78.
24 B-395 Pro Operation Manual, Version C
5.4.1.2 Electrode
8
4
765
1
2
3
4
Figure 5-8: Electrode with connecting elements
5 Putting into operation
a Screw nut M10 (polyamide)
O-Ring (10.82×1.78)
Insulator
Electrode
Connecting piece
5
Screw M4×12
6
The electrode is attached with either the elongated ring showing downwards or upwards so that
the distance between the nozzle and the electrode can be varied. The short distance between the
nozzle and the electrode is recommended during the production of small beads and if solutions of low
viscosity are used. The long distance is recommended during the production of large beads (approximately > 800 µm). The separation of the bead from the liquid jet should happen inside of the ring of
the electrode, where the electrostatic field is highest, or secondarily, in the space between the electrode and the nozzle, depending on the properties of your material.
5.4.1.3 Bead bypass system
The bead bypass system is used at the beginning and end of the encapsulation run to eliminate
unwanted beads produced by an unstable stream.
31 2
a Cup
Knob
Screw M5×8
Screw M3×6
Side axis
Main axis
Clip
O-Ring (6.1×1.6)
Figure 5-9: Bead bypass system - exploded view (left), assembled view (right)
25 B-395 Pro Operation Manual, Version C
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