Biorep Technologies PERI-4.2 Manual de usuario

PERIFUSION SYSTEM

MODEL No: PERI-4.2

______________________________________________________

USER MANUAL

______________________________________________________

TABLE OF CONTENTS

1GETTING STARTED .................................................................................................................................................................3

1.1 SYMBOLS USED IN THIS MANUAL...........................................................................................................................................................3

1.2 MACHINE INFORMATION.........................................................................................................................................................................3

1.3 CONTACT INFORMATION.........................................................................................................................................................................3

1.4 SAFETY INFORMATION............................................................................................................................................................................4

1.5 SITE PREPARATION .................................................................................................................................................................................5

1.6 PACKING LIST .........................................................................................................................................................................................5

2INTRODUCTION........................................................................................................................................................................6

3MACHINE PARTS IDENTIFICATION..................................................................................................................................7

4ACCESSORIES............................................................................................................................................................................9

4.1 TUBING SET ............................................................................................................................................................................................9

4.2 PERIFUSION CHAMBER............................................................................................................................................................................9

4.3 PERIFUSION CHAMBER AUXILIARY RACK ..............................................................................................................................................10

4.4 PERIFUSION NOZZLES ...........................................................................................................................................................................10

4.5 COLLECTION WELL-PLATES...................................................................................................................................................................10

4.6 BIO-GEL P-4.........................................................................................................................................................................................10

4.7 FIBERGLASS PRE-FILTERS......................................................................................................................................................................10

5PERIFUSION SYSTEM OVERVIEW....................................................................................................................................11

5.1 BASIC PERIFUSION SCHEMATIC.............................................................................................................................................................11

5.2 VALVE MANIFOLD................................................................................................................................................................................12

5.3 PERISTALTIC PUMP................................................................................................................................................................................14

5.4 PERIFUSION CHAMBER ..........................................................................................................................................................................14

5.5 COLLECTION TRAY................................................................................................................................................................................15

6PERIFUSION SYSTEM BASICS............................................................................................................................................16

6.1 POWER-UP............................................................................................................................................................................................16

6.2 LOGGING IN..........................................................................................................................................................................................17

6.3 MAIN MENU..........................................................................................................................................................................................18

6.4 TEST PAGE ............................................................................................................................................................................................19

6.5 SETTINGS ..............................................................................................................................................................................................21

7PERIFUSION OPERATION (PROTOCOL WIZARD) ......................................................................................................22

7.1 CHAMBER POSITIONING.........................................................................................................................................................................22

7.2 PROTOCOL.............................................................................................................................................................................................22

7.3 SYSTEM SETUP ......................................................................................................................................................................................23

7.4 CHAMBER SETUP ..................................................................................................................................................................................24

7.5 PERIFUSION TUBING SETUP...................................................................................................................................................................24

7.6 PRIMING................................................................................................................................................................................................25

7.7 CELL LOADING......................................................................................................................................................................................25

7.8 CHAMBER POSITIONING.........................................................................................................................................................................26

7.9 TEMPERATURE CONTROL ......................................................................................................................................................................26

7.10 PROTOCOL EXECUTION..........................................................................................................................................................................27

7.11 PROTOCOL EXAMPLES ..........................................................................................................................................................................29

8FINALIZING THE EXPERIMENT........................................................................................................................................31

8.1 COLLECTING THE CELLS........................................................................................................................................................................31

8.2 CLEANING THE SYSTEM.........................................................................................................................................................................32

9REPORT......................................................................................................................................................................................32

10 CUSTOMER SERVICE............................................................................................................................................................33

1 Getting Started

1.1 Symbols Used in this Manual

The lightning flash with arrowhead symbol, within an equilateral triangle, is intended to alert the user to

the presence of dangerous voltage within the machine’s enclosure that may be of sufficient magnitude

to constitute a risk of electric shock.

The exclamation point within an equilateral triangle is intended to alert the user to the presence of

important information in the literature that accompanies the device.

This symbol warns of moving parts that may cause personal injury.

This symbol is used when two people are needed to do the task.

1.2 Machine Information

In the spaces provided below, record the Model and Serial No. located on the rear panel of your machine.

Model No. __________________ Serial No.______________________

RETAIN THIS INFORMATION FOR FUTURE REFERENCE.

1.3 Contact Information

Biorep Technologies, Inc.

15804 NW 57th Avenue

Miami Lakes, FL 33014

[email protected]

www.biorep.com

Tel: 305-330-4449

Fax: 305-330-4402

WARNING:

Please, read this section before unpacking, installing or

operating this machine.

1.4 Safety Information

PLEASE READ AND OBSERVE ALL WARNINGS AND INSTRUCTIONS GIVEN IN THIS USER’S

MANUAL AND THOSE MARKED ON THE UNIT.

RETAIN THIS BOOKLET FOR FUTURE REFERENCE.

DO NOT REMOVE THE MACHINE’S COVER OR YOU MAY BE EXPOSED TO DANGEROUS

VOLTAGE. REFER SERVICING TO QUALIFIED PERSONNEL ONLY.

TO REDUCE THE RISK OF FIRE OR ELECTRIC SHOCK, DO NOT EXPOSE THIS DEVICE TO RAIN

OR MOISTURE. DANGEROUS HIGH VOLTAGES ARE PRESENT INSIDE THE ENCLOSURE.

READ AND UNDERSTAND HOW MOVING PARTS ARE CONTROLLED BEFORE OPERATING THE

MACHINE. KEEP YOUR FINGERS AWAY FROM MOVING PARTS; NEGLECTING TO DO SO MAY

CAUSE PERSONAL INJURY.

READ AND FOLLOW THESE INSTRUCTIONS:

1. Keep these instructions for future reference and heed all warnings stated in this manual.

2. Do not block any ventilation openings. Install the machine in accordance with the manufacturer’s

instructions. See Site Preparation in section 1.5 for details.

3. Do not defeat the safety purpose of the polarized or grounding-type plug. A polarized plug has two blades

with one wider than the other. A grounding-type plug has two blades and a third grounding prong, which

is provided for your safety. If the provided plug does not fit into your outlet, consult an electrician for

replacement of the obsolete outlet or contact Biorep Technologies, Inc. at (305) 330-4449.

4. Protect the power cord, the power entry module, and the plug from being walked on or pinched to avoid

damaging them.

5. Refer all servicing of the machine to qualified personnel. Servicing is required when the apparatus has

been damaged in any way, for example: the power-supply cord or plug is damaged, liquid has been

spilled or objects have fallen into the apparatus, or if the apparatus has been exposed to rain or moisture,

has been dropped, or does not operate normally for any reason.

WARNING:

DO NOT SKIP READING THIS SECTION.

1.5 Site Preparation

This machine is intended to work in a controlled laboratory environment. In order to properly place the machine,

please refer to the figure below. The machine has a ventilation opening in the rear, which should not be blocked.

A 12.7cm (5.0 in) clearance is recommended between the back of the machine and any obstacle. Bench-space

required is 49cm (19.3 in) x 65cm (25.6 in).

Figure 1: Site Preparation

1.6 Packing List

The following items are included with the purchase of a Perifusion system:

Qty

Reference

Description

86610610

Power Cord

PERI-CASSETTE

Pump cassettes

PERI-CHAMBER

Perifusion Chambers

PERI-TUB-12

Two-Stop Tygon Tubing Set (pack of 12)

PERI-TUB-040

Silicone Tubing (50 ft)

PERI-FILTER

Fiberglass Pre-filters (pack of 10)

PERI-BEADS-20

Bio-Gel P-4 Gel (20 grams)

PERI-RACK

Perifusion Chamber Rack

12885T21

Single Hole Punch

PERI-96-PLATE

96 well-plate

PERI-NOZZLE

Dispensing Nozzle

TUB-16

Size 16 Tubing for cooling tray

PERI-ORING

Perifusion Chamber O-rings

PERI-CLIPS

Pack of Clips

Table 1: Packing List

2 Introduction

Congratulations on your purchase of the Biorep Perifusion system. This is the newest generation of a product that

has received countless hours of development to include all the features necessary to perform the most controlled

and precise stimulated cell secretion experiments in the field. You will not be disappointed with the value and

efficiency this machine will bring to your research.

The Perifusion system accurately stimulates and collects the secretions of pancreatic islets in-vitro. Its use has

been extended to include the measurement of secretions of differentiated cells derived from pancreatic progenitor

stem cells. Coupled to a tandem mass spectrometer, the Perifusion system can help identify molecules secreted

during the stimulation of pancreatic islets.

The machine accurately controls temperature, flow, stimulus source and sample collection in up to 8 channels in

parallel. The Perifusion system is designed to execute pre-programmed protocols that can last for hours and

require several sample trays.

This User Manual is intended to be used by scientists, researchers, and technicians who have received training in

cell perifusion methods and technologies. This document contains the necessary information for installing and

operating the machine. However, it is not meant to provide scientific guidance on perifusion protocols. Any

perifusion protocols encountered in this manual will be solely for the purpose of illustration of machine operation

and should not be interpreted otherwise.

3 Perfusion

5 Purification

6 Perifusion

7 Cell Transport

8 Transplant

1 Donnor

2 Organ Transportation

4 Isolation

Figure 2 Perifusion System in the islet isolation process

3 Machine parts identification

Figure 3: Perifusion system, side view

Figure 4: Perifusion system, front view

Figure 5: Perifusion system, back view

1. Removable source holder

2. 32 (8 x 4) valve manifold for source handling

3. 12 Channel peristaltic pump (Main pump)

4. Temperature control inlet/outlets (x6)

5. Perifusion chamber

6. Sample collection tray

7. Alarm buzzer

8. Tray cooling diaphragm pump

9. Power cable

10. Incubator cover

11. 12” Touch-Screen

12. E-Stop

13. Orbitor Enable (Optional Add-On)

14. Nameplate

15. USB 2.0 Port 1

16. USB 2.0 Port 2

17. RJ45 LAN Port

18. Power Entry Module

4 Accessories

A complete set of accessories is included with the machine. For ordering additional or replacement accessories

please consult our webpage www.biorep.com or contact one of our sales representatives at (305) 330-4449.

4.1 Tubing Set

The Perifusion tubing set delivers the stimulus to the cell chamber while minimizing dead volume. The perifusion

tubing set is made out of 2 types of tubing: A- Connection tubing, which is for the perifusion fluid handling system

(source-manifold, manifold-pump, pump-chamber); and B- the peristaltic pump tubing, which provides the precise

resolution needed in perifusion experiments. A set of each type is included with this machine.

Item

Description

Reference

Silicone Tubing; 0.040in ID, 0.085in OD

89068-462

Two-Stop Tubing (Tygon R-3603) (orange-green);

0.015in ID, 0.035in WT;

PERI-TUB-12

Table 2: Items Included in the Perifusion Tubing Set

4.2 Perifusion Chamber

The Perifusion chamber is where all cell reactions will take place. Substances will be delivered through the inlet

tube, cells contained in the chamber will react to the substances, and secretions will come out the outlet tube for

collection. A new system includes 16 perifusion chambers.

1- Inlet tube

2- Top cap

3- Top O-Ring

4- Cell chamber

5- Bottom O-Ring + Filter

6- Bottom cap

7- Outlet tube

Figure 6: Perifusion Chamber

4.3 Perifusion chamber auxiliary rack

The auxiliary rack is included with every Perifusion system

to assist in the preparation of the Perifusion chambers. The

rack can hold up to 8 chambers at a time for cell loading

outside the machine. This allows the scientist to do the

preparation away from the machine in a more comfortable

position.

Figure 7: Perifusion Chamber Rack

4.4 Perifusion Nozzles

The Biorep® Perifusion Nozzle reduces the droplet size dispensed into the 96-well

plates. Small droplets (~4µL) ensure optimal resolution of the dynamic insulin

response.

Figure 8: Perifusion Nozzles

4.5 Collection well-plates

The collection well-plate is where the cell secretions and

the perfusate are collected for analysis. The Perifusion

system was designed to work with standard 96 well-plates,

and now has the ability to use high capacity, deep well

plates for longer experiments.

Figure 9: Collection Well-plate

4.6 Bio-Gel P-4

The Bio-Gel P-4 polyacrylamide beads are used at the time

of Perifusion chamber setup. They are used contain the

cells and prevent clogging of the filter.

See appendix A for beads solution preparation.

Figure 10: Bio-Rad polyacrylamide beads

4.7 Fiberglass pre-filters

Tabla de contenidos

Otros manuales de Equipo de laboratorio de Biorep Technologies

Biorep Technologies

Biorep Technologies PER-5.2 Manual de usuario

Manuales populares de Equipo de laboratorio de otras marcas

Qiagen

Qiagen DML 3000 Manual de usuario

Queensgate

Queensgate NANOSCAN OP400 Manual de programación

Conviron

Conviron GEN1000-GE Manual de usuario

Parr Instrument

Parr Instrument 4560 Manual de instrucciones

IKA

IKA IC control Manual de usuario

Integra

Integra DOSE IT Manual de usuario

Agilent Technologies

Agilent Technologies 5800 ICP-OES Manual de usuario

Endress+Hauser

Endress+Hauser Cleanfit CPA875 Manual de usuario

NI PXI-5422 Manual

Collomix

Collomix Aqix Manual de usuario

SPEX SamplePrep

SPEX SamplePrep 6875 Freezer/Mill Series Manual de usuario

Ocean Insight

Ocean Insight FLAME-NIR+ Manual de usuario

Parker

Parker ALIGN-MG-NA Manual de usuario

BD 644787 Manual de usuario

DENTAURUM

DENTAURUM Compact Megaplus Manual de usuario

Biuged Laboratory Instruments

Biuged Laboratory Instruments BGD 626 Manual de usuario

VWR

VWR SAS Super IAQ Manual de usuario

illumina

illumina MiSeqDx Manual de usuario