Eppendorf BioPhotometer plus User Manual

B 6132 900.017-00/1107

BioPhotometer plus

Operating manual

Trademarks

eppendorf and UVette are registered trademarks of Eppendorf AG, Hamburg, Germany. Cy is a registered trademark of GE Healthcare UK Ltd., Buckinghamshire, UK. Alexa Fluor is a registered trademark of Molecular Probes Inc., Eugene OR, USA. LabelGuard is a trademark of Implen GmbH, München, Germany.

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

6132 900.017-02/032012

345

Table of contents

BioPhotometer plus — Operating manual

Table of contents

1 User instructions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1.1 Using this manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1.2 Warning signs and hazard icons. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1.3 Symbols used. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

1.4 Abbreviations used. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7

2 Product description. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

2.1 Main illustration. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

2.2 Delivery package . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

2.3 Features . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 8

3 Safety . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

3.1 Intended use. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

3.2 Warnings for intended use . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 10

3.3 Application limits. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

3.4 Note on product liability . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 12

4 Installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

4.1 Preparing installation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

4.2 Selecting location . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

4.3 Connect device to the main power supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

4.4 Cuvettes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 13

4.5 Connect printer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14

Operating manual

5 Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

5.1 Overview of operating controls. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 15

5.2 Methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 16

5.3 Summary of the measuring procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

5.3.1 Prepare measurement . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

5.3.2 Select the method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 17

5.3.3 Measure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18

5.3.4 Finalize the method. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

5.4 Nucleic acids . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 19

5.5 Proteins. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

5.5.1 Protein 280 nm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

5.5.2 Protein after adding reagent (Bradford, BCA, Lowry) . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 20

5.6 Methods with evaluation via standards. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 21

5.7 OD 600 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

5.8 Dye-labeled biomolecules ("dye methods") . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

5.8.1 Method group "dye 550" . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

5.8.2 Method group "dye 650" . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 22

5.8.3 Frequency of incorporation "FOI" . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

5.8.4 Correction factors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

5.8.5 Measuring procedure and result display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 23

5.9 Methods for 340, 405 and 490 nm . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

5.10 Dilution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 24

5.11 Sample number . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 25

6 Parameter and functions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

6.1 Parameter. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

6.1.1 View, change and store the parameters of a method . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

6.1.2 Summary and description of parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 26

6.1.3 Parameters preprogrammed ex factory. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 28

6.2 Functions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 31

Operating manual

BioPhotometer plus — Operating manual

Table of contents

7 Maintenance . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

7.1 Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

7.1.1 Cleaning the cuvette shaft cover. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 32

7.2 Disinfection / Decontamination . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

7.3 Decontaminating before shipping . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

7.4 Replacing fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 33

7.5 Check photometer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 34

7.5.1 Test procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 35

8 Troubleshooting. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

8.1 Result flags. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 36

8.2 Error messages . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 37

8.3 General errors . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 38

9 Transport, storage and disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

9.1 Transport . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

9.2 Storage. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

9.3 Disposal . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39

10 Technical data . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

10.1 Power supply . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

10.2 Ambient conditions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

10.3 Weight / dimensions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

10.4 Interfaces . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 40

10.5 Photometer. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

10.6 Other technical parameters . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 41

10.7 Application parameters. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 42

11 Evaluation procedure . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

11.1 Evaluation with factor . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

11.2 Evaluation using standards. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

11.2.1 Single point calibration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 43

11.2.2 Multi-point calibration: calibration line . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

11.2.3 Multi-point calibration: Calibration curve . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

11.3 Dilution . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44

11.4 Special evaluation procedures for the dye methods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

11.4.1 Calculating the factor for the dye from the absorbance coefficient . . . . . . . . . . . . . . . . . . . . . . . . . . 45

11.4.2 Calculation of the FOI . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 45

11.5 Special evaluation procedures for nucleic acids and protein UV . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

11.5.1 Correction A

11.5.2 Correction A

11.5.3 Conversion into molar concentrations and nucleic acid amounts . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

12 Ordering information. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 48

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

340

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 46

550/650

BioPhotometer plus — Operating manual

1 User instructions

1.1 Using this manual



Before using the device for the first time, please read this operating manual.

 Please view this operating manual as part of the product and keep it somewhere easily

accessible.

 If this manual is lost, please request another one. The current version can be found on our

website, www.eppendorf.com (International) or www.eppendorfna.com (North America).

1.2 Warning signs and hazard icons

Depiction Meaning

DANGER

Risk of electric shock with potential for severe injury or death as a consequence.

DANGER

Risk of explosion with potential for severe injury or death as a consequence.

Operating manual

1.3 Symbols used

Depiction Meaning



•

(example)

Text Terms used in the device display.

DANGER

Biohazard with potential for risk to health or death as a consequence.

WARNING

Warning of potential injury or health risk.

CAUTION

Refers to risk of damage to property. Refers to particularly useful information and tips.

You are requested to perform an action. Perform these actions in the sequence described.

List. Press this key to perform the action described.

1.4 Abbreviations used

DNA Deoxyribonucleic acid

dsDNA double stranded DNA

Dye methods Group of methods via the keys dye 550 and dye 650

A Absorbance

FOI Frequency of Incorporation: measure for the number of dye molecules related to the number of

nucleotides in dye-labeled biomolecules

M mol/l (molar)

OD600 Optical density for wave length 600 nm

RNA Ribonucleic acid

ssDNA Single stranded DNA

UV Ultraviolet radiation VK Coefficient of variation (standard deviation / mean), in percentages

Operating manual

BioPhotometer plus — Operating manual

2 Product description

2.1 Main illustration

Abb. 1: Front and rear view

Fig. 1: Front and rear view

1 Device display 2 Cuvette shaft cover

Slide back or forward to open or close.

3 Cuvette shaft 4 Connection RS-232

5 ID plate 6 Mains connection socket

7 Fuse holder 8 Mains switch

9 Measuring keys 10 Keyboard

2.2 Delivery package

Number Description

1 BioPhotometer plus 1 Power cable 2 UVette

Original Eppendorf plastic cuvette, individually wrapped, for direct use in the BioPhotometer, certified RNase, DNA and protein free

1 BioPhotometer plus operating instructions, multilingual,

2.3 Features

Cuvette photometer The BioPhotometer plus is a cuvette photometer for the fast, simple and comfortable

measurement of the most important methods in the molecular biology and biochemistry research laboratory. It can also be used for the main photometric methods in cell biology.

Method programs Method programs for calculating the concentration of nucleic acids, proteins and dye-labeled

nucleic acids and proteins as well as the method "OD600" for calculating the bacteria density through measuring turbidity are already preprogrammed. However, you can modify those in many parameters. Other methods for calculating the concentration for 340, 405 and 490 nm can be freely programmed. The method "absorbance" is used for the fast absorbance measurement with any of 9 available wavelengths without further evaluation.

Method programs are combined into groups which you can open quickly via fixed keys.

Cuvettes You can use standard rectangular glass or plastic cuvettes with optical transparency for the

respective measuring wavelength. With the Eppendorf UVette you can also measure nucleic acids and proteins in the UV range using a plastic cuvette. A cover protects the cuvette shaft against dust and other contamination if the photometer is not in use. To open the cuvette shaft it is moved back, to cover it after completing the measurements it is moved forward.

Measuring keys After opening a method the device is immediately ready for measuring. A measurement is started

with one of the 3 round measuring keys.

BioPhotometer plus — Operating manual

2 Product description

Evaluation The BioPhotometer plus converts the measured absorbance values into concentration results.

Dependent on the method the results can be calculated via fixed factors, standards, or curve calibration. In addition to the results the device also displays the absorbance values and some other important details, e.g. the common absorbance quotients for nucleic acid calculations. Sample dilutions can also be included in the evaluation. Other special evaluation procedures are provided for specific method groups. For example, when calculating the concentration of dyed nucleic acids the frequency of incorporation related to the amount of nucleic acid can also be calculated.

Output The BioPhotometer plus outputs the results via the device display and via a printer available from

Eppendorf.

Operating manual

BioPhotometer plus — Operating manual

3 Safety

3Safety

3.1 Intended use

The intended area of use for the BioPhotometer plus is the research laboratory in molecular biology, biochemistry and cell biology. The device may only be operated by trained specialist staff.  The BioPhotometer plus is used to perform photometric measurements to quantify biomolecules as well as to perform turbidity measurements of microbiological cultures in routine laboratories. Due to the specific examination of selected parameters, the device serves to monitor laboratory processes. Use only Eppendorf accessories or accessories recommended by Eppendorf AG.

3.2 Warnings for intended use

Operating manual

DANGER!

Danger! Electric shock from damage to device/power cable.

 Only switch on the device if the device and the power cable are undamaged.  Only use devices that have been properly installed or repaired.

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

 Switch off the device and disconnect it from the power supply before starting cleaning or

disinfecting.

 Do not allow any liquids to penetrate the inside of the housing.  Do not disinfect by means of spraying.  Only reconnect the device to the power supply once it is completely dry.

Danger! Electric shock.

 Switch off the device and disconnect the power plug before opening the device to replace the

fuses. These tasks may only be performed by appropriately trained staff.

Risk of explosion!

 Do not operate the device in rooms where work is being carried out with explosive

substances.

 Do not use this device to process any explosive, radioactive or highly reactive substances.  Do not use this device to process any substances, which could create an explosive

atmosphere.

DANGER!

WARNING!

Risk when handling toxic or radioactively-marked liquids or pathogenic germs.

 Follow national regulations governing the handling of these substances.  For complete instructions regarding the handling of germs or biological material of risk group

II or higher, please refer to the "Laboratory Biosafety Manual" (Source: World Health Organization, current edition of the Laboratory Biosafety Manual).

Warning! Damage to health from chemicals.

Hazardous chemicals cause burns and other health hazards.

 Follow the instructions for use provided by the manufacturers of reagents and other

chemicals.

Warning! Poor safety due to incorrect accessories.

The use of accessories and spare parts other than those recommended by Eppendorf may impair the safety, function and precision of the device. Eppendorf accepts no warranty or liability for damage caused by third-party parts or incorrect use.

 Use only original accessories recommended by Eppendorf.

3 Safety

WARNING!

BioPhotometer plus — Operating manual

Warning! Risk to health from contaminated device

 Perform decontamination before storing or dispatching the device and/or its accessories.

Operating manual

CAUTION!

Caution when using aggressive chemicals.

Aggressive chemicals may damage both the device and its accessories.

 Do not use any aggressive chemicals on the device or its accessories, such as strong and

weak alkalis, strong and weak acids, acetone, formaldehyde, chlorinated hydrocarbons or phenol.

 If the device becomes contaminated with aggressive chemicals, clean it immediately with a

neutral cleaning agent.

Caution! Corrosion from 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 prolonged

periods.

Caution! Damage to electronic components from condensation.

 After moving the device from a cooler environment (e.g., cool room or outdoors), wait at least

an hour before connecting it to the mains power supply.

Caution! Function may be impaired by mechanical damage.

 After a mechancial damage to the device ensure by means of an inspection that the

measuring and evaluation functions of the device function correctly.

Caution! Damage due to overheating.

 Do not place the device close to sources of heat (e.g., radiator, drying cabinet).  Do not expose the device to direct sunlight.  Allow air to circulate freely by leaving at least 5 cm to adjoining devices or to the wall and

keep the underside of the device free.

CAUTION!

Caution! Material damage from incorrect use.

 Only use the product for its intended purpose as described in the operating manual.  Ensure adequate material resistance when using chemical substances.  In cases of doubt, contact the product manufacturer.

Caution! Poor safety due to missing operating manual.

 When passing on the device, always enclose the operating manual.  If you lose the operating manual, request a replacement. The current version of the

operationg manual and the safety instructions can also be found on our website www.eppendorf.com.

Caution! Damage as a result of incorrect packing.

Eppendorf accepts no warranty or liability for damage caused by incorrect packing.

 Only dispatch the device in the original packaging provided for carriage.

BioPhotometer plus — Operating manual

3 Safety

Operating manual

CAUTION!

Caution! Damage from improper cleaning of the cuvette shaft.

 Only clean the cuvette shaft using a moist cotton swab.(see Cleaning on page 32)  Do not allow any liquid to enter the cuvette shaft.  Do not reach with your fingers into the cuvette shaft.

CAUTION!

Caution! Faulty measurement due to device confusion.

 If you use the devices Biophotometer 6131 and BioPhotometer plus 6132 in your laboratory,

note the different method designations on the keys.

3.3 Application limits

Risk of explosion!

 Do not operate the device in rooms where work is being carried out with explosive

DANGER!

substances.

 Do not use this device to process any explosive, radioactive or highly reactive substances.  Do not use this device to process any substances, which could create an explosive

atmosphere.

3.4 Note on product liability

In the following cases, the protection provided in the device may be impaired: Liability for the function of the device passes to the operator if:

• the device is not used in accordance with the operating manual.

• the device is used outside the sphere of application described here.

• the device is used with accessories and consumables (e.g. tubes and plates), which are not

recommended by Eppendorf AG.

• the device is maintained or repaired by persons not authorized by Eppendorf.

• the owner has made unauthorized modifications to the device.

4 Installation

4.1 Preparing installation



Keep the transport carton and the packing material for subsequent safe transport or storage.

 Check the completeness of delivery based on the details of the scope of delivery (see

Delivery package on page 8).

 Check all parts for any transport damage.

4.2 Selecting location

Select the location for the BioPhotometer plus in accordance with the following criteria:

• 2 power sockets with ground conductor for the BioPhotometer plus and the printer.

• Solid laboratory bench with horizontal work surface

Space requirement of the device: 40 cm (with printer: 65 cm) width, 50 cm depth.

• Temperature: 15 to 35 °C. Avoid direct sunlight.

• Humidity: 25 to 75 % relative humidity.

• Atmospheric pressure: 70 to 106 kPa.

BioPhotometer plus — Operating manual

Operating manual

4.3 Connect device to the main power supply

1. Place the BioPhotometer plus onto a suitable work surface.

2. Ensure that the mains voltage and frequency match the details for the range of mains voltages and frequencies on the device nameplate.

3. Connect the device to the power supply and switch it on from the mains power switch 8 (Fig. 1 on p. 8).

4. Remove the protective film from the device display.

4.4 Cuvettes

You can insert standard rectangular glass or plastic cuvettes into the cuvette shaft (outside diameter 12.5 mm x 12.5 mm). The optical path height must be 8.5 mm above the cuvette base and the total cuvette height must be at least 36 mm. The light beam in the cuvette is 1.0 mm wide and 1.5 mm high.

The cuvettes must be optically transparent for the respective measuring wavelength. For measurements in the UV range Eppendorf provides a plastic cuvette called UVette which is transparent from wavelengths above 220 nm and therefore also suitable for the measurement of nucleic acids.

Abb. 2: Overview of different cuvette types

Fig. 2: Overview of different cuvette types

Operating manual

BioPhotometer plus — Operating manual

4 Installation

4.5 Connect printer

You can connect the Eppendorf thermal printer to the serial interface RS-232 C of the photometer (see Ordering information on page 48).

1. Connect the printer cable to the serial printer port 4 of the photometer and tighten the locking screws.

2. Connect the printer cable to the printer and also tighten the locking screws.

3. Connect the printer to the power supply using the plug-in power unit supplied (printer accessory) and switch it on.

4. Check the printer settings in accordance with the following table and make corrections where necessary.

Information about modifying printer settings can be found in the operating manual for the printer.

Tab. 1: Setting the DIP SW for the thermal printer

DIP SW-1 Meaning

1 (OFF) Input = Serial 2 (ON) Printing Speed = High 3 (ON) Auto Loading = ON 4 (OFF) Auto LF = OFF 5 (ON) Setting Command = Enable 6 (OFF) Printing 7 (ON) Density 8 (ON) = 100%

DIP SW-2 Meaning

1 (ON) Printing Columns = 40 2 (ON) User Font Back-up = ON 3 (ON) Character Select = Normal 4 (ON) Zero = Normal 5 (ON) International 6 (ON) Character 7 (ON) Set 8 (OFF) = U.S.A.

DIP SW-3 Meaning

1 (ON) Data Length = 8 bits 2 (ON) Parity Setting = NO 3 (ON) Parity Condition = Odd 4 (OFF) Busy Control = XON/XOFF 5 (OFF) Baud 6 (ON) Rate 7 (ON) Select 8 (ON) = 9600 bps

BioPhotometer plus — Operating manual

5 Operation

5.1 Overview of operating controls

Abb. 3: Control panel of the BioPhotometer plus.

Fig. 3: Control panel of the BioPhotometer plus.

Operating manual

Key Function Oval blue keys, e.g.:

• In the method selection: select method group.

• When entering values: enter digits.

Circular keys

Start standard measurement.

Start blank measurement.

Start sample measurement.

Oval transparent keys

• In the method selection: open parameter list.

• During the measurement procedure: enter sample dilution.

• In the method selection: open function list.

• During the measuring procedure: modify sample number.

• When entering digits: enter decimal point.

• Confirm entry or selection.

• Open selected method or function.

Cursor keys

Delete entry

Move cursor up or down in the device display.

Operating manual

BioPhotometer plus — Operating manual

5 Operation

5.2 Methods

The following methods are available and already preprogrammed ex factory. You can modify most parameters and save them as a modified method (see Parameter on page 26).

Method group Method Explanation Wavelength

DNA dsDNA Calculating the concentration of DNA with

ssDNA OLIGO DNA

RNA RNA Analogous to method group DNA. As method group DNA.

OLIGO RNA

Protein BCA Calculating the concentration of proteins after

BCA MICRO BRADFORD 595 nm BRADFORD MICRO LOWRY 595 nm LOWRY MICRO PROTEIN 280 nm Calculating the concentration of proteins with

OD600 OD600 Turbidity measurement to determine the

dye 550 DYE 550-dsDNA For dye-labeled biomolecules: calculating the

DYE 550-ssDNA DYE 550-RNA DYE 550-OLIGO DYE 550-PROTEIN

dye 650 DYE 650-dsDNA Analogous to method group "dye 550". DNA/RNA/OLIGO: see method

DYE 650-ssDNA DYE 650-RNA DYE 650-OLIGO DYE 650-PROTEIN

assay 340 ASSAY 340/1 Calculating the concentration by measuring at

ASSAY 340/2 ASSAY 340/3

assay 405 ASSAY 405/1 Analogous to method group "assay 340". 405 nm

ASSAY 405/2 ASSAY 405/3

assay 490 ASSAY 490/1 Analogous to method group "assay 340". 490 nm

ASSAY 490/2 ASSAY 490/3

ABSORBANCE ABSORBANCE Rapid absorbance measurement after

evaluation via factor. The methods differ mainly in the preprogrammed factor.

adding reagent. The methods are preprogrammed with the evaluation procedure calibration curve. Number and target concentration of the standards can be modified.

evaluation via factor.

bacteria density.

concentration of the molecule (nucleic acid or protein) and the dye in a single measuring procedure. The frequency of incorporation of the dye in the biomolecule is also determined.

340 nm. The evaluation procedures can be freely programmed. As a sample the methods are already preprogrammed with the following evaluation procedures: 340/1: evaluation via factor 340/2: evaluation via a standard. 340/3: evaluation via calibration curve with 6 standards.

selecting the wavelength.

Measuring wavelength: 260 nm Secondary wavelengths to check for purity: 230, 280, 340 nm

550 nm

Measuring wavelength: 280 nm Secondary wavelengths to check for purity: 260, 340 nm

595 nm

DNA/RNA/OLIGO: see method groups DNA and RNA PROTEIN: see method PROTEIN 280 nm Measuring wavelength for the dye: 550 nm

groups DNA and RNA PROTEIN: see method PROTEIN 280 nm Measuring wavelength for the dye: 650 nm

340 nm

230, 260, 280, 340, 405, 490, 550, 595, 650 nm

+ 35 hidden pages