Eppendorf AF2200 User Manual

PlateReadergN)manualAF2200

Eppendorf® PlateReader AF2200

Operating manual

Copyright© 2013 Eppendorf AG, Hamburg. No part of this publication may be reproduced without the prior permission of the copyright owner.

Trademarks

Eppendorf® and the Eppendorf Logo are registered trademarks of Eppendorf AG, Hamburg, Germany.

Microsoft®, Windows® and Excel® are registered trademarks of Microsoft Corporation, Redmond, WA, USA.

Pentium® is a registered trademark of Intel Corporation, Santa Clara, CA, USA.

Greiner® is a registered trademark of Greiner Bio-One GmbH, Frickenhausen, Germany.

Trademarks are not marked in all cases with ™ or ® in this manual.

6141 900.208-02/072013

Table of contents

Eppendorf® PlateReader AF2200 3

English (EN)

Table of contents

1 Operating instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.2 Danger symbols and danger levels . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.2.1 Danger symbols . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7 1.2.2 Danger levels. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1.3 Symbols used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8 1.4 Abbreviations used . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

2 Product description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11

2.1 Main illustration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 2.2 Delivery package. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 11 2.3 Features. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 2.3.1 Methods and templates. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12 2.3.2 Result output . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

2.4 System requirements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13 2.5 Fluorescence detection system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15 2.5.1 Light source system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16 2.5.2 Upper fluorescence optics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17 2.5.3 Lower fluorescence optics . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18 2.5.4 Fluorescence detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

2.6 Absorbance measurement system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19 2.6.1 Light source system . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 2.6.2 Absorbance optics. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20 2.6.3 Absorbance detection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

3 Safety. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

3.1 Intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 3.2 User profile . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 3.3 Warnings for intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 3.3.1 Personal injury . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21 3.3.2 Damage to device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

3.4 Information on product liability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

4 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

4.1 Preparing installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 4.2 Removing the transport locks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25 4.3 Selecting the location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 4.4 Connecting the device to the mains . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27 4.5 Connecting the device to a computer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 27

5 Inserting the filter slide . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

5.1 Information on optical filters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 5.1.1 Fluorescence filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29 5.1.2 Absorbance filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 29

5.2 Filter slides and filter orientation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 5.2.1 Filter slides . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30 5.2.2 Inserting optical filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30

Table of contents

4Eppendorf® PlateReader AF2200 English (EN)

5.3 Installing user-specific filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 5.3.1 Removing filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31 5.3.2 Inserting user-specific filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32 5.3.3 Defining filters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

6 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

6.1 Overview of operating controls . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 6.1.1 Notes on operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 6.1.2 Switching on the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 6.1.3 Starting the device . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37 6.1.4 Disconnecting the device from the computer and switching it off . . . . . . . . . . . . . . . . 37 6.1.5 Loading the device with plates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

6.2 Settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 6.2.1 Temperature control . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 6.2.2 Kinetic measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 6.2.3 Shaking microplates . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38 6.2.4 Multi mode measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 6.2.5 Ratio mode measurements (= measurements well by well) . . . . . . . . . . . . . . . . . . . . . 39

6.3 Optimizing fluorescence measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 6.3.1 Gain settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 6.3.2 PMT properties . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39 6.3.3 Flash settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 6.3.4 Settle time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 6.3.5 Recommended filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40 6.3.6 Recommended microplate types. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

6.4 Optimizing absorbance measurements. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 6.4.1 Flash settings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 6.4.2 Settle time . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 6.4.3 Multiple reads per well . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41 6.4.4 MRW type . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42 6.4.5 MRW size . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 6.4.6 MRW limit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43 6.4.7 Displaying results in Microsoft Excel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44 6.4.8 Various MRW software functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

7 Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

7.1 General errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45 7.2 Error messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

8 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47

8.1 Decontamination before shipment . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 8.2 Checking air filters for contamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 47 8.3 Replacing air filters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 8.4 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48 8.5 Disinfection/Decontamination. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49 8.6 Regular quality checks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 49

8.6.1 Test criteria . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 50 8.6.2 Test instructions, upper fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 51 8.6.3 Test instructions, lower fluorescence . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 55 8.6.4 Absorbance test instructions - baseline stability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 60

Table of contents

Eppendorf® PlateReader AF2200 5

English (EN)

9 Technical data. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61

9.1 Power supply. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 9.2 Weight/dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 9.3 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 9.4 Measuring methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 9.4.1 Notes on measuring methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 61 9.5 Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62

9.6 Fluorescence intensity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 62 9.6.1 Definition of the detection limit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 9.6.2 Upper fluorescein (fluorescence intensity) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63 9.6.3 Lower fluorescein (fluorescence intensity) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 63

9.7 Absorbance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64 9.8 "Quick" measurements . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 64

10 Ordering Information . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 65

11 Transport, storage and disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67

11.1 Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 11.2 Inserting transport locks . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 67 11.3 Storage . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68 11.4 Disposal. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 68

Table of contents

6Eppendorf® PlateReader AF2200 English (EN)

Operating instructions	7
Eppendorf® PlateReader AF2200
English (EN)

1Operating instructions

1.1Using this manual

Read this operating manual completely before using the device for the first time. Please also note the operating instructions for the accessories, if applicable.

This operating manual is part of the product. Thus, it must always be easily accessible.

Enclose this operating manual when transferring the device to third parties.

If this manual is lost, please request another one. For the current version, please refer to our webpage www.eppendorf.com/worldwide (international) or www.eppendorfna.com (North America).

1.2Danger symbols and danger levels

The safety instructions in this manual appear with the following danger symbols and danger levels:

1.2.1	Danger symbols
	Biohazard	Explosion
	Electric shock	Toxic substances
	Hazard point	Material damage

1.2.2Danger levels

DANGER	Will lead to severe injuries or death.
WARNING	Can lead to severe injuries or death.
CAUTION	Can lead to light to moderate injuries.
ATTENTION	May lead to material damage.

Operating instructions

8Eppendorf® PlateReader AF2200 English (EN)

1.3Symbols used

Depiction			Meaning
1.			Actions in the specified order
2.
			Actions without a specified order
•			List:
Text			Display or software texts
			Additional information

Operating instructions	9
Eppendorf® PlateReader AF2200
English (EN)

1.4Abbreviations used

A

Absorbance

ANSI

American National Standards Institute

DNA

Deoxyribonucleic acid (DNA)

dsDNA

Double-stranded DNA

M

mol/L (molar)

MTP

Micro test plate

OD600

Optical density at a wavelength of 600 nm

RFU

Relative Fluorescence Unit: measure for the intensity during fluorescence measurements

RNA

Ribonucleic acid (RNA)

SBS

Society for Biomolecular Screening

ssDNA

Single-stranded DNA

USB

Universal Serial Bus

UV

Ultraviolet radiation

Vis

Visible light

CV

Coefficient of variation (standard deviation/average value) in percent

Operating instructions

10Eppendorf® PlateReader AF2200 English (EN)

Product description

Eppendorf® PlateReader AF2200 11

English (EN)

2Product description

2.1Main illustration

7

1
	2	3	4	5	6
Fig. 2-1:	Front and rear view

1	Filter slide flap	5	USB port, USB connector Typ B
2	Button for moving the plate compartment in	6	Warranty seal
	and out	7	Name plate
3	Plate compartment flap
4	Mains connection socket with power switch
	0 = switched off, I = switched on.

The device may only be opened by the manufacturer's service technicians. The warranty is void if the warranty seal has been removed or damaged.

2.2Delivery package

Quantity Description

1PlateReader AF2200

1Mains and USB cable

1Hardware manual

1CD with software and software manual

Product description

12Eppendorf® PlateReader AF2200 English (EN)

2.3Features

The PlateReader AF2200 is well-suited for the quantification of biomolecules, such as nucleic acids and proteins, and for measuring fluorescence-based assays in plate format. A wide variety of methods can be executed in the UV/Vis range, and various fluorescence dyes can be detected.

The PlateReader AF2200 is designed for use in molecular biology, biochemistry and cell biology research laboratories. The device was developed for use as a universal laboratory device for processing biological and non-biological samples. 6-well to 384-well microplates which comply with ANSI/SLAS standards can be used.

Additional features:

•Dual-mode plate reader for measuring UV/Vis absorption and upper and lower fluorescence intensity.

•Pre-configured and freely configurable filter combinations.

•Preprogrammed methods for a quick start.

•Simple data evaluation (factor or standard-based) in Excel format.

•Clearly designed software with multi-user license.

•Optional μPlate for measuring the smallest sample volumes with your own methods.

•Heatable measurement chamber; shaking mode that allows for linear and orbital shaking of the micro plate.

2.3.1Methods and templates

Numerous methods for the concentration determination of nucleic acids and proteins (direct and dye-labeled) are preprogrammed. Evaluations are carried out via factor or standard curve. Furthermore, templates are already preprogrammed for various measuring procedures (e.g., kinetic methods). For these templates, only the raw data is provided via Excel.

2.3.2Result output

The PlateReader AF2200 uses Excel documents to output raw data and results. These files can be saved and processed as needed.

Product description	13
Eppendorf® PlateReader AF2200
English (EN)

2.4System requirements

To install the software, the system must meet the following minimum requirements:

Component	Minimum	Recommended
Computer	Windows XP (32-bit):	2 GHz (dual-core)
	Windows-compatible computer with a
	Pentium-compatible processor, 1 GHz
	Windows Vista (32-bit):	2 GHz (dual-core)
	Windows-compatible computer with a
	Pentium-compatible processor, 1 GHz
	Windows 7 (32or 64-bit):	2 GHz (dual-core)
	Windows-compatible computer with a
	Pentium-compatible processor, 1 GHz
	Windows 8 (32or 64-bit):	2 GHz (dual-core)
	Windows-compatible computer with a
	Pentium-compatible processor, 1 GHz
Operating system	Windows XP (32-bit) SP3	Windows 7 (64-bit) SP1
	Windows Vista (32-bit)
	Windows 7 (32-bit)
	Windows 7 (64-bit)
	Windows 8 (32-bit)
	Windows 8 (64-bit)
Memory	Windows XP: 512 MB RAM	1 GB RAM
	Windows Vista (32-bit): 1 GB RAM	2 GB RAM
	Windows 7 (32-bit): 1 GB RAM	2 GB RAM
	Windows 7 (64-bit): 2 GB RAM	3 GB RAM
	Windows 8 (32-bit): 1 GB RAM	2 GB RAM
	Windows 8 (64-bit): 2 GB RAM	3 GB RAM
Memory requirements	700 MB RAM	1 GB RAM
Monitor	Super VGA Graphics	1280 x 1024
Color depth	256	-
Mouse	Microsoft-compatible mouse or similar	-
	device
Communication	1x USB 2.0	2x USB 2.0
		1x RS232 (serial)

Product description

14Eppendorf® PlateReader AF2200 English (EN)

Component	Minimum	Recommended
Drive, graphic card	1xCD-ROM drive	-
	Windows Vista:
	DirectX 9 graphic and 32 MB graphics
	memory (for Home Basic version)
	DirectX 9 graphic and 128 MB graphics
	memory with WDDM support for all
	other versions.
	Windows 7:
	DirectX 9 graphic with WDDM driver
	1.0 or higher.
.NET	Microsoft .NET Framework 2.0	-
	If this version is not available, it will be
	installed by the installation and update
	program using any available installation
	of the .NET Framework.
	Under Windows 8, Microsoft .NET
	Framework 3.5 (incl. 2.0) has to be
	installed by the user.
Windows Installer	3.1	-
	If this version is not available, it will be
	installed by the installation and update
	program.
Microsoft Excel	Excel 2002 (only compatible for	-
	script-based messages without data
	reduction). Excel 2002 is not
	compatible for methods.
	Excel 2003, 2007, 2010 (32-bit) and
	2013 (32-bit)
	The Starter Edition is not supported.

To support new versions of Windows, there will be future software updates for the PlateReader AF2200. Please contact your local Eppendorf contact person if you have any questions.

Product description	15
Eppendorf® PlateReader AF2200
English (EN)

2.5Fluorescence detection system

reference diode

condensor

flash

lamp

1

PMT

3

emission

filter

beam

splitter

excitation	2
filter

2

MTP

Fig. 2-2: Upper fluorescence detection system (fluorescence top reading)

Product description

16Eppendorf® PlateReader AF2200 English (EN)

reference diode

condensor

flash

lamp

1

PMT

	3
		bottom
	emission	emission
	filter	fiber
	mirror
excitation	2
filter

MTP

mirror

bottom excitation fiber

2

Fig. 2-3: Lower fluorescence detection system (fluorescence bottom reading)

1 Light source

3 Fluorescence detector

2 Fluorescence optics

2.5.1Light source system

Flash light enters the optical system focused by the aperture with the excitation filter. The opening functions as a color-specific light source.

The light source system consists of the following components:

•Flash lamp

•Condenser optics

•Excitation filter

•Flash lamp monitor (= reference diode)

Product description

Eppendorf® PlateReader AF2200 17

English (EN)

Flash lamp

The device uses a high-performance xenon gas-discharge lamp (flash lamp). The flash jumps over a small gap between two electrodes. The lamp contains high-pressure xenon. The flash disappears within several microseconds. The flash frequency is 40 Hz.

The device uses the flash lamp for fluorescence and absorbance measurements. The primary advantages of this unique type of lamp are:

•High intensity from the deep UV range through near IR range

•Very long service life

•A single light source for a wide range of applications

•No heat-up time required

Condenser

A condenser is used to focus the light into the optical fluorescence system via the entrance aperture.

Excitation filter

For fluorescence applications, optical band-pass filters are required which allow suitable wavelengths to pass through and filter the rest of the spectrum.

In many cases, the fluorescence emissions spectra are not dependent on the precise excitation wavelength. Therefore, a relatively wide band-pass filter (10 – 40 nm) can be used for a maximum total fluorescence signal. The filters are mounted in removable slides.

Flash monitoring

The light energy of individual flashes may fluctuate slightly. To take these fluctuations into account, a silicon photodiode monitors the energy of each individual flash. The results of the fluorescence measurement will be compensated accordingly.

2.5.2Upper fluorescence optics

Flash light enters the optical system focused through an aperture. The opening functions as a color-specific light source. A partially transparent mirror (beam splitter) reflects 50 % of the light toward the microplate. The lens system of the objective focuses the light on the sample.

The fluorescence emission in the well will be measured from above. The fluorescence light is bundled by the lens, guided through the partially transparent mirror with emission filter and focused via the exit aperture so that it can be detected.

Product description

18Eppendorf® PlateReader AF2200 English (EN)

The lens system

The lens system bundles the maximum amount of fluorescent light from a well and focuses it onto the detection system through the exit aperture.

The lenses are made of quartz glass. This material is extremely UV-transparent and virtually non-autofluorescent.

2.5.3Lower fluorescence optics

Flash light enters the optical system focused through an aperture. The opening functions as a color-specific light source. The lower excitation fiber guides the light onto the lower measuring head, which consists of an elliptical mirror. The mirror focuses the light into the well through the base of the microplate.

The emitted light will be focused onto the lower emission fiber which guides the light into the fluorescence detection system via a mirror and through the emission filter.

2.5.4Fluorescence detection

Emission filter

Emission filters are used to differentiate between non-specific fluorescence signals and sample-specific fluorescence signals. The filters are mounted in removable slides. The spot diameter of the light beam on the filters is approx. 2 mm for the upper and 2 mm for the lower optical system.

PMT detection

A photomultiplier is used for detection as the light intensities of the fluorescence can be very low.

Product description

Eppendorf® PlateReader AF2200 19

English (EN)

2.6Absorbance measurement system

A light path similar to the one used for fluorescence excitation is implemented for absorbance measurements. The absorbance measurement module is located below the plate compartment. It measures the light that is transmitted through the sample. A reference measurement, during which the plate compartment is positioned outside of the light beam, is carried out before the microplate is measured.

reference

condensor

diode

beam

splitter

flash

lamp

1

absorbance

2

filter

2

MTP

		absorbance		3
		photo diode
Fig. 2-4: Absorbance measurement system
1	Light source		3 Absorbance detector
2	Absorbance optics

Product description

20Eppendorf® PlateReader AF2200 English (EN)

2.6.1Light source system

The absorbance light source system is similar to the upper fluorescence system (see p. 16).

Absorbance filter

For absorbance applications, optical band-pass filters are required to enable suitable wavelengths to pass through and to filter the rest of the flash lamp spectrum. The filters are mounted in removable slides.

For absorbance measurements, filters with relatively narrow bandwidths (2 – 10 nm) and steep slopes are required.

2.6.2Absorbance optics

The mirror carriage has an absorbance position. Two small apertures create a light beam which is narrow and more strongly collimated than with fluorescence excitation.

Light that is focussed through the dispensed liquid will be slightly refracted at the transitions between air, liquid and the plate bottom. To achieve a reliable measurement even with a sample that is not flat, a convex lens bundles the light beams, which may have been deflected too far from the optical axis.

The spot diameter of the absorbance light beam is 0.5 mm.

2.6.3Absorbance detection

Use the Eppendorf μPlate G0.5 for absorbance measurements with smaller volumes (2 μL).

A silicon photodiode is used to measure the light beam. It is sensitive across a wide range of wavelengths and is suitable for light intensities that occur with absorbance measurements up to 4 OD.

Safety

Eppendorf® PlateReader AF2200 21

English (EN)

3Safety

3.1Intended use

The PlateReader AF2200 is designed for use in molecular biology, biochemistry and cell biology research laboratories. The device was developed for use as a universal laboratory device for processing biological and non-biological samples.

The PlateReader AF2200 is intended for indoor use only. All country-specific safety requirements for operating electrical equipment in the laboratory must be observed.

The PlateReader AF2200 is used for photometric concentration determination of analytes in liquids and for measuring fluorescence-based assays. Only 6-well to 384-well microplates that comply with ANSI/SLAS standards can be used.

Use Eppendorf accessories or accessories recommended by Eppendorf only.

3.2User profile

The device and accessories may only be operated by trained and skilled personnel.

Before using the device, read the operating manual carefully and familiarize yourself with the device's mode of operation.

3.3Warnings for intended use

3.3.1Personal injury

DANGER! Electric shock as a result of penetration of liquid.

Switch off the device and disconnect the power plug before starting cleaning or disinfection work.

Do not allow any liquids to penetrate the inside of the housing.

Do not spray clean/spray disinfect the housing.

Only plug the device back in if it is completely dry, both inside and outside.

WARNING! Electric shock due to damage to device or mains cable.

Only switch on the device if the device and mains cable are undamaged.

Only use devices that have been properly installed or repaired.

In case of danger, disconnect the device from the mains supply by pulling the power plug from the device or the mains socket or, by using the isolating device intended for this purpose (e.g. emergency stop switch in the laboratory).

Safety

22Eppendorf® PlateReader AF2200 English (EN)

WARNING! Damage to health from toxic, radioactive or aggressive chemicals as well as infectious liquids and pathogenic germs.

Observe the national regulations for handling these substances, the biological security level of your laboratory, the material safety data sheets and the manufacturer's application notes.

Wear personal protective equipment.

CAUTION! Poor safety due to incorrect accessories and spare parts.

The use of accessories and spare parts other than those recommended by Eppendorf may impair the safety, functioning and precision of the device. Eppendorf cannot be held liable or accept any liability for damage resulting from the use of incorrect or non-recommended accessories and spare parts, or from the improper use of such equipment.

Only use accessories and original spare parts recommended by Eppendorf.

WARNING! Damage to health due to contaminated device and accessories.

Decontaminate the device and the accessories before storage and shipping.

3.3.2Damage to device

NOTICE! Damage from the use of aggressive chemicals.

Do not use any aggressive chemicals on the device or its accessories, such as strong and weak bases, strong acids, acetone, formaldehyde, halogenated hydrocarbons or phenol.

If the device has been contaminated by aggressive chemicals, clean it immediately using a mild cleaning agent.

NOTICE! Corrosion due to aggressive cleaning agents and disinfectants.

Do not use corrosive cleaning agents, aggressive solvents or abrasive polishes.

Do not incubate the accessories in aggressive cleaning agents or disinfectants for longer periods.

NOTICE! Damage to electronic components due to condensation.

Condensate can form in the device after it has been moved from a cool environment to a warmer environment.

After installing the device, wait for at least 3 h. Only then connect the device to the mains.