Bruker ImagePrep Bedienungsanleitung

Revision 4 (April 2013)

ImagePrep

User Manual

(for Software Version 2.0)

Page 2 of 77 ImagePrep User Manual Revision 4

Bruker Daltonik GmbH

Reproduction, adaptation, or

translation without prior written

permission is prohibited, except

as allowed under the copyright

laws.

Document History

See Table of Changes

First edition: Aug 2007

Printed in Germany

Warranty

The information contained in this

document is subject to change

without notice.

Bruker Daltonik GmbH makes no

warranty of any kind with regard

to this material, including, but not

limited to, the implied warranties

of merchantability and fitness for

a particular purpose.

Bruker Daltonik GmbH shall not

be liable for errors contained

herein or for incidental or

consequential damages in

connec

tion with the furnishing,

performance or use of this

material.

Bruker Daltonik GmbH assumes

no responsibility for the use or

reliability of its software on

equipment that is not furnished

by Bruker Daltonik GmbH.

Safety Information

Safety class

The ins

trument is a Safety

Class I instrument and has

been designed and tested in

accordance with IEC

Publication 1010: Safety

Requirements for Electrical

Equipment for Measurement,

Control, and Laboratory Use.

WARNING

Connecting an instrument to a

power source

which has not

equipped with a protective

earth contact creates a shock

hazard for the operator and can

damage the instrument.

Likewise, interrupting the

protective conductor inside or

outside the instrument or

disconnecting the protective

earth terminal c

reates a shock

hazard for the operator and can

damage the instrument.

WARNING

The instrument has to be

disconnected from its power

source before the inner cover is

removed or

the instrument is

otherwise opened

All connections of the

instrument have to be used in

correct way.

It is only permitted to use wires

and cables provided

by the

manufacturer.

Instrument Identification

Each instrument is identified by

a unique serial number. This

serial numbers is located on a

label at

the rear of the

instrument.

When corresponding with

Bruker Daltonik GmbH about

your instrument, be sure to

include the full serial number.

Write the serial number of your

instrument here for reference:

Serial #:

Manual Conventions

Cautions

Cautions call attention to

procedures which, if not correctly

performed or adhered to, could

result in

damage to the

instrument.

Warnings

Warnings call attention to

procedures which, if not correctly

performed or adhered to, could

result in personal injury.

Part Numbers

In this manual

, Bruker Daltonik

GmbH part numbers are generally

listed in parentheses after the

name of the part or in tables in the

parts section.

A few tools and supplies listed

have no part numbers and are not

available from Bruker Daltonik

GmbH. Most of these can be

obtained from laboratory supply

companies.

ImagePrep User Manual

# 8261654

ImagePrep User Manual, Revision 4 (April 2013)

Fahrenheitstr. 4

D-28359 Bremen

Germany

Phone: +49 (421) 2205-345

FAX: +49 (421) 2205-103

Email: maldi.sw.support@bdal.de

Internet: http://www.bruker.com

Bruker Daltonik GmbH Safety Symbols

ImagePrep User Manual Revision 4 Page 3 of 77

Safety Symbols

The following symbols appear in this manual where special attention is necessary.

NOTE

This symbol is used to draw attention to points that may affect

the operation or performance of the equipment.

WARNING

This symbol is used to indicate processes and procedures that

involve harmful substances ( e.g. Organic solvents).

Take appropriate care and refer to the Material Safety data Sheets

for the substances.

Equipment Disposal

Under Europe

an Directive 2002/96/EC, this label applies to electrical

and electronic equipment installed and sold in the European Union.

The consumer is committed by law to dispose of such equipment at the

end of its lifetime. When disposing of the equipment or parts thereof, it

must not be disposed of in household waste. To do so can contaminate

and pollute the environment.

To dispose of the equipment responsibly, contact your community

waste disposal authority. The details are regulated in national law.

Technical Support Bruker Daltonik GmbH

Page 4 of 77 ImagePrep User Manual Revision 4

Technical Support

If you need any assistance with this product please call or write to:

Address: BRUKER Daltonik GmbH,

Fahrenheitstr. 4,

28359 Bremen,

Germany

Phone: +49 (0)421 2205-345 Technical / Software Support

+49 (0)421 2205-200 Sales Department

Fax: +49 (0)421 2205-103 Technical / Software Support

E-mail: maldi.sw.support@bdal.de Technical / Software Support

sales@bdal.de Sales Department

Internet: http://www.bruker.com

Table of Changes

Revision

Date Changes Remarks

1 Aug 2007 First edition -

1.1 Oct 2007 Editorial update -

2 Aug 2009 Updated hardware and ImagePrep software

Version 2.2 -

3 Jun 2010 Procedural changes

4 Apr 2013 WEEE, SAP renumbering and web address

update, added figure and table numbering

Bruker Daltonik GmbH Getting Started

ImagePrep User Manual Revision 4 Page 5 of 77

Getting Started

•Read the manual carefully!

•Sections written in black letters in the Table of Contents contain important

information for all users.

•Sections written in orange (light-grey) letters in the Table of Contents contain

important information for expert users.

•Place the instrument in a ventilated fume hood.

•Connect the instrument nitrogen source with a pressure with about

to 2-8 bar.

•Set the local date and time (see section 6.7).

•The instrument was delivered with a pre-adjusted spray head.

Prior to first use check the spray performance with methanol as described in

section 6.2.

What’s New in Version 2.0

•New spray head is supported section 6.1

•Simplified Spray Head Adjustment procedure section 6.2

•Improved sample loading procedure section 3.3

•New Test Spray buttons section 3.5.3

•Improved clean method section 3.5.5

•Diverse bug fixes

Table of Contents Bruker Daltonik GmbH

Page 6 of 77 ImagePrep User Manual Revision 4

Table of Contents

all users / experts

COPYRIGHT .................................................................................................................II

SAFETY SYMBOLS........................................................................................................III

TECHNICAL SUPPORT..................................................................................................IV

Table of Changes.................................................................................................. iv

GETTING STARTED.......................................................................................................V

TABLE OF CONTENTS..................................................................................................VI

1SAFETY AND SITE REQUIREMENT ISSUES................................................................8

1.1 Safety Precautions........................................................................................8

1.2 Installation and Site Requirements................................................................9

1.3 List of Approved Chemicals ..........................................................................9

2GENERAL INTRODUCTION ....................................................................................10

3SAMPLE PREPARATION WORKFLOW DESCRIPTION ...............................................12

3.1 Switching-on / Rebooting............................................................................12

3.2 Preparing the Tissue Sections ....................................................................13

3.3 Loading the Sample....................................................................................14

3.4 Loading the Matrix Solution.........................................................................15

3.5 Automatic Sample Preparation....................................................................16

3.5.1 Select Method........................................................................................17

3.5.2 Customize Method .................................................................................19

3.5.3 Test Spray and Start Automatic Preparation...........................................21

3.5.4 Mounting the Glass Slide into the Adapter..............................................22

3.5.5 Cleaning the Spray Chamber and the Spray Head.................................23

3.6 Manual Sample Preparation with ImagePrep..............................................25

Bruker Daltonik GmbH Table of Contents

ImagePrep User Manual Revision 4 Page 7 of 77

all users / experts

4 Method Development.......................................................................................26

4.1 Hardware and Software Requirements....................................................26

4.2 Interpretation of the Scattered Light Curves ..........................................27

4.3 The Method Development GUI.................................................................29

4.3.1 Method Access window.......................................................................29

4.3.2 Starting Methods for Test Purposes...................................................31

4.3.3 Method Editor: Phase Overview Level................................................32

4.3.4 Editing a Phase.....................................................................................35

4.3.4.1 The Matrix Thickness Window......................................................36

4.3.4.2 The Nebulize Window....................................................................39

4.3.4.3 The Incubation Window ................................................................45

4.3.4.4 The Dry Window ............................................................................47

4.4 General Method Development Rules.......................................................50

5TROUBLESHOOTING ............................................................................................52

5.1 Error Messages ..........................................................................................52

5.2 Other Problems...........................................................................................55

5.3 FAQ (Frequently Asked Questions) ............................................................59

6MAINTENANCE ....................................................................................................62

6.1 Spray Generator Replacement ...................................................................62

6.2 Global Spray Power Adjustment .................................................................64

6.3 Dismount the Spray Head...........................................................................66

6.4 Installation of Software and Method Updates..............................................66

6.5 Generate a Status Report...........................................................................67

6.6 System Reboot (see section 3.1)................................................................69

6.7 Date / Time Setting.....................................................................................69

6.8 Touch Panel Stylus.....................................................................................69

6.9 Filter Replacement......................................................................................69

6.10 Fuse Replacement......................................................................................69

6.11 List of Spare Parts ......................................................................................70

Appendix 1:.............................................................................................................71

Appendix 2:.............................................................................................................73

Bruker Daltonik GmbH Safety and Site Requirement Issues

Page 8 of 77 ImagePrep User Manual Revision 4

1 Safety and Site Requirement Issues

1.1 Safety Precautions

WARNING

Operate instrument

in ventilated fume hood only

This instrument vaporizes chemicals

and emits organic vapors.

NOTE

Only use manufacturer’s approved chemicals!

See list in section 1.3

NOTE

Never disable or bypass any of the safety features.

Figure

Instrument in

ventilated

fume hood

Bruker Daltonik GmbH Safety and Site Requirement Issues

ImagePrep User Manual Revision 4 Page 9 of 77

1.2 Installation and Site Requirements

•Fume hood mandatory

•Power 110-240 VAC,

50/60 Hz

200 VA

•Nitrogen - Quality 4.0 (99.99%)

or better

- 2-8 bar (29 – 116 PSI)

- 4 mm tubing

(or 1/8’’ with adapter)

•Temperature 18-24°C (64–75°F)

for best analytical performance

For details please refer to Site Preparation

Specifications document.

1.3 List of Approved Chemicals

•Select the solvents from the list

(or mixtures thereof) (see Figure 3).

•Cleaning solvent: methanol.

Do not use other solvents for cleaning!

•All common MALDI matrices can be used.

The matrix concentration should be at least 20%

below saturation.

•Optionally you can acidify the solution with up to

0.5% TFA.

NOTE

Other solvents or chemicals can be used with

manufacturer’s agreement.

Water

Ethanol

Methanol

2-Propanol

Acetonitrile

Figure

2 Connections

Figure

List of

solvents

General Introduction Bruker Daltonik GmbH

Page 10 of 77 ImagePrep User Manual Revision 4

2 General Introduction

Sample preparation is crucial for the quality of MALDI-tissue-imaging data.

Unfortunately, the current matrix application protocols have significant

disadvantages:

While pneumatic spray preparations provide good homogeneity and spatial

resolution of the images, the process is manual and highly irreproducible.

Depending of the degree of tissue wetting either the analyte molecules are badly

incorporated into the matrix (too dry) or the spatial resolution is lost (too wet).

Nano-spotting on the other hand provides quality spectra but as a sequential

process it is slow, spatial resolution is limited by the spot raster (typical >200 µm)

and perfect alignment with the mass spectrometer is critical.

ImagePrep is a new preparation device for MALDI Imaging that reproducibly

provides high resolution images at high speed and high spectra quality at the same

time.

On tissue, a lateral resolution of 50 µm can be achieved.

In the ImagePrep device, matrix

aerosol is created by vibrational

vaporization (see Figure 4), that is

gently deposited onto tissue

sections. During the whole

preparation process the spray

chamber is filled with nitrogen in

order to prevent sample oxidation

and to obtain reproducible

experimental conditions

independent of the humidity in the

ambient air.

An average droplet size of 20μm is generated with all droplet diameters being

smaller than 50μm, which corresponds well to the final matrix crystal size.

Multiple sections on a microscope slide can be homogeneously matrix-coated,

typically with 30-100 cycles within about one hour. Each cycle consists of three

phases:

1. deposit droplet layer,

2. incubate in controlled atmosphere, and

3. allow partial/complete drying.

Figure

4 Vibrational vaporization

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