Molecular Devices SPECTRAmax PLUS 384 Operator's Manual

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SPECTRAmax

PLUS

384

Microplate Spectrophotometer Operator’s Manual

Molecular Devices Corporation

Part # 0112-0056 Rev. A

1311 Orleans Drive Sunnyvale, California 94089

Molecular Devices Corporation SPECTRAmax

PLUS

384

Operator’s Manual



Copyright 2000, Molecular Devices Corporation. All rights reserved. No part of this publication may be reproduced, transmitted, transcribed, stored in a retrieval system, or translated into any language or computer language, in any form or by any means, electronic, mechanical, magnetic, optical, chemical, manual, or otherwise, without the prior written permission of Molecular Devices Corporation, 1311 Orleans Drive, Sunnyvale, California, 94089, United States of America.

Patents

The SPECTRAmax PLUS U.S. Patents

4,968,148 and 5,112,134 ; Canadian Patent 1,305,334 .

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and methods are protected by the following patents:

Other U.S. and International patents pending.

Trademarks

SPECTRAplate, SPECTRAstrip, PathCheck and Automix are trademarks and SPECTRAmax and SOFTmax are registered trademarks of Molecular Devices Corporation.

Apple and Macintosh are registered trademarks of Apple Computer, Incorporated. All other company and product names are trademarks or registered trademarks of their respective owners.

Disclaimer

Molecular Devices Corporation reserves the right to change its products and services at any time to incorporate technological developments. This manual is subject to change without notice.

Although this manual has been prepared with every precaution to ensure accuracy, Molecular Devices Corporation assumes no liability for any errors or omissions, nor for any damages resulting from the application or use of this information.

Molecular Devices Corporation Instrument Warranty

Molecular Devices Corporation warrants this product against defects in material or workmanship as follows:

1) All parts of the SPECTRAmax

PLUS

Microplate Spectrophotometer are warranted for a period of one (1)

384

year from the original date of delivery.

2) All labor charges to repair the product for a period of one (1) year from the original date of delivery will be paid by Molecular Devices Corporation.

3) This warranty covers the SPECTRAmax printer, software, reagents, or disposables used with the instrument.

Labor and Parts

To obtain warranty service during the applicable warranty period, you must take the product or deliver the product properly packaged in the original shipping materials and carton to an authorized Molecular Devices Corporation service facility. You must call or write to the nearest Molecular Devices Corporation service facility to schedule warranty service. You may call Molecular Devices Corporation at the telephone number or address below to locate the nearest service facility. You must schedule warranty service time of requesting warranty service, you must present proof of purchase documentation which includes the date of purchase and Molecular Devices Corporation must have the enclosed Warranty Registration form completed, signed and returned by you within ten (10) working days of the date of delivery.

ii SPECTRAmax PLUS

384

PLUS

instrument only, and does not extend to any computer,

prior to bringing or shipping the product for servicing. At the

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Microplate Spectrophotometer Operator’s Manual

This warranty only covers defects arising under normal usage and does not cover malfunctions of failures resulting from misuse, abuse, neglect, alteration, modification, or repairs by other than an authorized Molecular Devices Corporation service facility.

Repair or replacement as provided under this warranty is the exclusive remedy of the purchaser (the “Buyer”).

Molecular Devices Corporation (the “Seller”) shall not be liable for any incidental or consequential damages for breach of any express or implied warranty on this product, except to the extent required by applicable law. The Seller specifically excludes all express and implied warranties including without limitation any implied warranty that the products sold under this agreement are merchantable or are fit for any particular purpose, except such warranties expressly identified as warranties and set forth in the Seller’s current operating manual, catalog or written guarantee covering such product. The Seller also makes no warranty that the products sold under this agreement are delivered free of the rightful claim of any third party by way of patent infringement or the like.

claim which arises out of compliance with the specifications. Any description of the products contained in this agreement is for the sole purpose of identifying them. Any such

description is not part of the basis of the bargain, and does not constitute a warranty that the products shall conform to that description. Any sample or model used in connection with this agreement is for illustrative purposes only, is not part of the basis of the bargain, and is not to be construed as a warranty that the products will conform to the sample or model. No affirmation of fact or promise made by the Seller, whether or not in this agreement, shall constitute a warranty that the products will conform to the affirmation or the promise.

For the name of the nearest authorized Molecular Devices Corporation service facility, please contact Molecular Devices at one of the following telephone numbers:

(408) 747-1700 (800) 635-5577 (U.S. and California)

If the Buyer furnishes specifications to the Seller, the Buyer agrees to hold the Seller harmless against any

Microplate Spectrophotometer

100-240V ~ 4A 50-60 HZ T4.0A

SPECTRAmax PLUS

2 Lines Fused, unplug before servicing!

Vor Wartungsarbeiten Netzstecker ziehen!

Sunnyvale, CAMade in USA

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Microplate Spectrophotometer Operator’s Manual

Contents

Figures and Table. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .vi

Conventions Used in this Manual . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii

Glossary of Terms . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . vii

Chapter 1 Instrument Description. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-1

Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3

General Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3

PathCheck

™. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4

Component Description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5

Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5

Control Panel Display . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-7

The Microplate Drawer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-8

The Cuvette Chamber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9

The Back Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-10

Functional Description . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-11

Stand-Alone Operation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-11

Wavelength Selection. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-11

Temperature Regulation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-11

Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-12

Printed Data Output . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-12

Computer Control. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-13

Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-16

Chapter 2 Installation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-1

Installation Warnings. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3

Installation Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3

Unpacking . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-3

Setting Up for Stand-Alone Use. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-4

Chapter 3 Stand-Alone Operation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-1

Prepare for a Reading . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Read the Cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Operation Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

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Microplate Spectrophotometer Operator’s Manual v

Turn the Instrument and Printer On . . . . . . . . . . . . . . . . . . . . . . 3-3

Set the Temperature. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Select the Wavelength . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Chapter 4 Maintenance and Troubleshooting . . . . . . . . . . . . . . . . . . . . . . . . . 4-1

Technical Support . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3

Warnings and Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3

General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4

Cleaning. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4

Changing the Fan Filter . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4

Changing the Fuses . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-5

Error Codes and Probable Causes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-7

Error Messages. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-7

Opening the Drawer Manually . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-11

Appendix A Printers and Cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-1

Compatible Printers . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3

Cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3

Appendix B Accessories. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-1

Items Available for Use with the SPECTRAmax PLUS

384

. . . . . . . . . . . . B-3

Figures

Table

Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . I-1

Figure 1.1: SPECTRAmax PLUS

Figure 1.2: Major Areas of the SPECTRAmax PLUS

384

. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3

384

. . . . . . . . . . . . 1-5

Figure 1.3: Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5

Figure 1.4: Microplate Drawer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-8

Figure 1.5: Cuvette Chamber . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9

Figure 1.6: Test Tube Cover . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9

Figure 1.7: Components on the Back Panel of the

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SPECTRAmax PLUS

. . . . . . . . . . . . . . . . . . . . . . . . . . 1-10

Figure 1.8: Sample Printout from Thermal Printer . . . . . . . . . . . . . . . 1-13

Figure 2.1: View of Rear Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2-4

Figure 4.1: Power Switch, Fuse Box, and Power Receptacle . . . . . . . . . 4-5

Figure 4.2: Removing the Fuse Box. . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-6

Figure 4.3: The Fuse Box and Holder (with Fuses) Removed . . . . . . . . 4-6

Table 4.1: Error Codes . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-8

vi SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Conventions Used in this Manual

Glossary of Terms

The names of keys that appear on the SPECTRAmax PLUS shown in boxed Helvetica type. Example:

[Drawer]

384

control panel are

Italic and boldface type are used for emphasis. Examples: “Press carefully to engage,” “

NOTE:

Do not press down. ”

A note provides information that will help you properly execute an

action or procedure.

CAUTION:

Indicates an action or condition that could potentially damage

the instrument or one of its components or could result in loss of data.

WARNING:

person working with the system

BIOHAZARD:

Indicates a situation that could result in potential injury to a

Indicates a condition involving potentially infectious biolog-

ical agents requiring that proper handling precautions be taken.

Absorbance, A

The amount of light absorbed by a solution. To measure absorbance accurately, it is necessary to eliminate light scatter. In the absence of turbidity, absorbance = optical density.

A = log (I

= incident light

/I)

I = transmitted light

In this manual, we use the terms absorbance and optical density interchangeably.

Optical Density, OD

The amount of light passing through a sample to a detector relative to the total amount of light available. Optical Density includes absorbance of the sample plus light scatter from turbidity.

Transmittance, T

The ratio of transmitted light to the incident light.

T = I/I %T = 100 T

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Microplate Spectrophotometer Operator’s Manual vii

viii SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Chapter 1 Instrument Description

Introduction. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3

General Overview . . . . . . . . . . . . . . . . . . . . . . . . 1-3

PathCheck

Component Description . . . . . . . . . . . . . . . . . . . . . . . . . 1-5

Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-5

Control Panel Display . . . . . . . . . . . . . . . . . . . . . 1-7

The Microplate Drawer . . . . . . . . . . . . . . . . . . . . 1-8

The Cuvette Chamber . . . . . . . . . . . . . . . . . . . . . 1-9

The Back Panel . . . . . . . . . . . . . . . . . . . . . . . . . 1-10

Functional Description . . . . . . . . . . . . . . . . . . . . . . . . . 1-11

Stand-Alone Operation . . . . . . . . . . . . . . . . . . . 1-11

Wavelength Selection. . . . . . . . . . . . . . . . . . . . . 1-11

Temperature Regulation . . . . . . . . . . . . . . . . . . 1-11

Data Collection . . . . . . . . . . . . . . . . . . . . . . . . . 1-12

Printed Data Output . . . . . . . . . . . . . . . . . . . . . 1-12

Computer Control. . . . . . . . . . . . . . . . . . . . . . . 1-13

Specifications . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-16

™

. . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-4

Chapter 1

1-2 SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

Introduction

General Overview

The SPECTRAmax™ PLUS and sensitive UV/VIS measurements of a variety of analytes including speciﬁc proteins, nucleic acids, and other molecules across a wide range of concentrations. It measures the optical density (OD) of samples in both 96- and 384-well microplates or in a cuvette at a selected wavelength for a single point in time (endpoint), over a speciﬁed period of time (kinetic), or over a selected wavelength range (spectral scan).

384

microplate spectrophotometer provides rapid

Figure 1.1 : The SPECTRAmax PLUS

384

System

The on-board microprocessor calculates and reports the absorbance (or % Transmittance) for each well of a microplate. Data from multiple wavelengths can be acquired during a single reading, if desired, and different calculations can be made based on this data using SOFTmax PRO software, including the subtraction of blanks, use of standard curves, etc.

Typical applications include endpoint assays (quantitation of cytoproliferation by MTT reduction, colorimetric protein assays, protein measurement at 280 nm, DNA/RNA measurement at 260 nm, and endpoint ELISAs) and kinetic measurements (enzyme studies, such as determination of the activity of enzymes released from cells, and kinetic ELISAs).

All standard 96- and 384-well microplates, strip wells, and ﬁlter-bottom microplates can be used in the SPECTRAmax

PLUS

384

. When reading at wavelengths below 340 nm, special UV-transparent, disposable or quartz microplates and cuvettes that allow transmission of the

far UV spectra must be used.

With SOFTmax PRO, the contents of the wells in a microplate can be mixed automatically by shaking before each read cycle, making it possible to perform kinetic analysis of solid-phase, enzyme-mediated reactions (mixing is not critical for liquid-phase reactions). The temperature of the microplate chamber can also be regulated, if desired, from 4°C above ambient to 45°C.

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Microplate Spectrophotometer Operator’s Manual 1-3

Chapter 1

An optional printer, for use with the cuvette chamber in a stand-alone mode, can be purchased as an accessory.

PathCheck

™

As predicted by the Beer Lambert law of light absorption, absorbance is proportional to the distance that light travels through the sample—the longer the pathlength, the higher the absorbance.

Microplate readers use a vertical light path so the distance of the light through the sample depends on the volume. This variable pathlength makes it difﬁcult to perform extinction-based assays and confusing to compare results between microplate readers and spectrophotometers.

Horizonta

The standard pathlength of a cuvette is the conventional basis for quantifying the unique absorptivity properties of compounds in solution.

Horizontal

Light Path

Quantitative analyses can be performed on the basis of extinction coefﬁcients, without standard curves (e.g., protein at 280 nm, nucleic acid estimates at 260 nm and NADH-based enzyme assays). When using a cuvette, the pathlength is known and is

independent of sample volume, so absorbance is

Cuvette

proportional to concentration.

dependent on the

both

Vertical

In the microplate, pathlength is liquid volume, so absorbance is proportional to the concentration and the pathlength of the sample. Standard curves are often used to determine analyte concentrations in vertical-beam photometry of

Vertical Light Path

unknowns, yet errors can still arise from pipetting the samples and standards. The PathCheck feature of the SPECTRAmax

PLUS

384

automatically determines the pathlength of aqueous samples in the microplate and normalizes the absorbance in each well to a pathlength of 1 cm. This novel approach to

Microplate Wells

correcting the microwell absorbance values is accurate to within 2.5% of the values obtained directly in a 1cm cuvette.

Reference measurements made by reading the cuvette or using factory-stored values derived from deionized water can be used to normalize the OD data for microplate wells.

This pathlength correction is accomplished only with the use of SOFTmax PRO software. Stand-alone control of the instrument is limited to ﬁxed-wavelength

1-4 SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

Component Description

Control Panel

reference measurements made with the cuvette. SOFTmax

PRO software from

Molecular Devices provides full instrument control and statistical data analysis.

The main components of the SPECTRAmax PLUS

384

are:

• The control panel

• The microplate drawer

• The cuvette chamber

• The back panel (connections and power switch)

Back Panel

Cuvette Chamber

Microplate Drawer

Figure 1.2 : Major areas of the SPECTRAmax PLUS

384

Control Panel

Figure 1.3 : Control Panel

The control panel consists of an LCD and nine pressure-sensitive, color-coded membrane keys which can be used to perform the stand-alone functionality of the SPECTRAmax plate drawer and cuvette chamber, setting the wavelength at which to read the cuvette, toggling between displayed values as absorbance or percent transmission,

PLUS

384

, including setting the temperature inside the micro-

SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual 1-5

Chapter 1

and opening or closing the microplate drawer. Pressing a control panel key activates the function.

LCD

A 2-

-20-character liquid crystal display which shows the current instrument set-

tings.

Keys

TEMP

on/off

(Incubator) Enables/disables the incubator function.

(Incubator) Allows you to enter a set point to regulate the

microplate and cuvette chamber temperature. Pressing the up or down arrow key scrolls up or down, starting at the previous

TEMP

temperature setting (or the default of 37.0°C, if no setting had

▲

been made). Pressing the up (

) or down (

▼

) arrow once incre-

ments or decrements the temperature shown in the display by

0.1°C; pressing and holding

either arrow increments or decrements the temperature shown in the display by 1°C until it is released. If you increment the setting to the highest limit (45°C) and continue to press the up (

▲

) arrow, the display will not change. If you decrement the setting to the lowest limit, 15°C, and continue to press the down (

▼

) arrow, the display will not

change.

CAUTION:

If the incubator is disabled, pressing the

will

enable the incubator.

[Temp On/Off]

key

Selects the wavelength that will be used for reading the cuvette manually. Pressing the up or down arrow key scrolls up or down through the available wavelengths, starting at the previ-

ous setting. Pressing the up (

▲

) or down (

▼

) arrow once increments or decrements the wavelength shown in the display by 1 nm; pressing and

holding either arrow increments or decre-

ments the wavelength shown in the display by 10 nm until it is released. If you increment the setting to the highest limit (1000 nm) and continue pressing the up (

▲

) arrow, the display returns to the lowest possible setting (190 nm) and begins incrementing from there. The inverse is true for decrementing by pressing the down (

▼

) arrow.

NOTE:

1-6 SPECTRAmax PLUS

Setting the wavelength in SOFTmax PRO and reading using the soft-

ware will override the manual setting.

The control panel will not show

the wavelength selected through SOFTmax PRO.

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

[REF]

READ

CUVETTE

[%T/A] [Drawer]

NOTE:

A reading of buffer, water, or air taken in the cuvette that is

used as I reference reading is taken, the SPECTRAmax

the I

to calculate Absorbance or %Transmittance. If no

384

PLUS

values stored in the NVRAM (non-volatile memory) of

will use

the instrument.

Initiates the sample reading of the cuvette.

Toggle switch used to display cuvette data as percent

transmission or absorbance.

Opens or closes (toggles) the microplate drawer. Whether or not the drawer will remain open depends on the incubator setting. If the incubator is off, the drawer will remain open; if the incubator is on, the drawer will close after approximately 10 seconds to assist in maintaining temperature control within the microplate chamber.

If the SPECTRAmax

software, the

[READ CUVETTE]

If the SPECTRAmax

PLUS

384

is being controlled by the computer

and

384

is idle, all keys on the control panel are

[REF]

button will be inactive.

active. During a kinetic run, if the read interval is >30 seconds, pressing the

[Drawer]

button will open the drawer. The instrument will automat-

ically close the microplate drawer for the next reading.

Control Panel Display

The left side of the display shows the temperature, both actual and set point, and whether or not the temperature is at the set point (enunciator blinks if not at set point). The middle of the display shows the wavelength. The right side of the display shows the data received from the reading as either absorbance or percent transmission and indicates whether or not a reference measurement was made (enunciator blinks if no reference reading was taken).

To change the contrast on the control panel, press or down keys.

[%T/A]

and the

[Temp]

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Microplate Spectrophotometer Operator’s Manual 1-7

Chapter 1

A1 Indicator

The Microplate Drawer

Figure 1.4 : Microplate Drawer

The microplate drawer is located on the right side of the SPECTRAmax PLUS

384

and slides in and out of the reading chamber. Springs on two sides of the drawer automatically position and hold a microplate in the proper position. The drawer remains in the reading chamber during read cycles.

Microplate drawer operation varies, depending upon the incubator status. To open the drawer, press the

[Drawer]

key. If the incubator is turned on, the drawer will remain open for approximately ten seconds, after which a beeping sound will alert you approximately two seconds before the drawer closes automatically.

NOTE:

Do not obstruct the movement of the drawer. If you must retrieve a plate after an error condition or power outage and the drawer will not open, it is possible to open it manually (see Chapter 4, “Maintenance and Troubleshooting”).

Microplates

The SPECTRAmax PLUS microplates, strip wells, and ﬁlter-bottom microplates. When reading at wavelengths below 340 nm, special UV-transparent, disposable or quartz microplates allowing transmission of the deep UV spectra must be used.

Not all manufacturers’ microplates are the same with regard to design, materials, or conﬁguration. Temperature uniformity within the microplate may vary depending on the type of microplate used.

384

can accommodate standard 96- and 384-well

1-8 SPECTRAmax PLUS

384

Microplate Spectrophotometer Operator’s Manual

The Cuvette Chamber

Instrument Description

Figure 1.5 : Cuvette Chamber

Located at the right front of the SPECTRAmax PLUS

384

, the cuvette chamber has a lid that lifts up, allowing you to insert or remove a cuvette. The chamber contains springs that automatically position the cuvette in the proper alignment for a reading. The cuvette door must be closed in order to initiate a reading.

Cuvettes

The SPECTRAmax PLUS cuvettes and 12 x 75 mm test tubes when used with the test tube cover.

Figure 1.6 : Test Tube Cover

Not all manufacturers’ cuvettes are the same with regard to design, materials, or conﬁguration. Temperature uniformity within the cuvette may vary depending on the type of cuvette used.

384

can accommodate standard-height (45 mm), 1-cm

Be sure to handle cuvettes on the frosted sides only. Place the cuvette into the chamber so that the “reading” (clear) sides face left and right (see Figure 1.5).

NOTE:

SPECTRAmax PLUS

384

Microplate Spectrophotometer Operator’s Manual 1-9

The SPECTRAmax

micro cuvettes.

PLUS

384

will not accept the short (25-mm high)

Chapter 1

The Back Panel

Power Switch

Fuse Box Cover

Label

Power Cord

Receptacle

Printer (Centronics)

Computer (RS-232)

Figure 1.7: Components on the Back Panel of the SPECTRAmax PLUS

384

The following components are located on the back panel of the SPECTRAmax

384

PLUS

system:

• Power switch—a rocker switch, labeled I/O (for on and off, respectively).

• Power cord receptacle—plug the power cord in here.

• Fuse box cover—cannot be opened while the power cord is plugged in. When

opened, it provides access to the fuse box containing two fuses that are required for operation.

• Printer port (double-shielded, 25-pin parallel, for use in stand-alone opera-

tion)—plug the 25-pin end of the cable into this port; the other (Centronics) end attaches to the port on the printer.

• Computer port (double-shielded 8-pin RS-232 serial, for use with an external

computer)—plug the 8-pin DIN serial cable connecter into this port; the other end attaches to the serial (modem) port of the computer.

• Labels—provide information about the SPECTRAmax PLUS

voltage rating, cautionary information, serial number, etc. Record the serial number shown on this label for use when contacting Molecular Devices Technical Services.

1-10 SPECTRAmax PLUS

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, such as line

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

Functional Description

Stand-Alone Operation

The paragraphs that follow describe functions that can be activated during standalone operation.

When operating the SPECTRAmax PLUS

384

as a stand-alone system, you can

obtain a ﬁxed-wavelength reading of the cuvette. The wavelength is chosen by

pressing the up or down arrows beneath the display labeled λ. Temperature regu-

lation of the cuvette/microplate chamber can be enabled and set using the keys on the left side of the control panel and actual temperature can be monitored in the display. To obtain a reading of the cuvette, press either the reference reading) or the

[READ CUVETTE] key (for a sample reading). The

[REF] key (for a

resulting data can be displayed as absorbance or percent transmission by toggling the

[%T/A] key. You can also open or close the drawer using the [Drawer] key

on the right of the control panel.

NOTE:

Keep cuvette door closed when reading a cuvette.

Wavelength Selection

The display on the control panel shows the currently selected measurement wave-

length (λ). You can change the wavelength by pressing the up or down arrows

until the display shows the desired wavelength. The wavelength will wrap around to the top or bottom when the last selectable wavelength is reached.

Temperature Regulation

The SPECTRAmax PLUS of the microplate chamber from 4°C above ambient to 45°C. Upon power up, when the incubator is off, the temperature in the SPECTRAmax PLUS plate chamber is ambient and isothermal. Pressing the incubator

key will cause the SPECTRAmax to begin warming the microplate/cuvette

Off]

chamber. The temperature set point defaults to 37.0°C at start-up.

NOTE:

Accuracy of the temperature set point is only guaranteed if the set point is at least 4°C above ambient. If the temperature set point is lower than the ambient temperature, the chamber temperature will remain at ambient. Temperature regulation is controlled by heaters only and, therefore, cannot cool the temperature to a setting lower than ambient. Additionally, the highest setting (45°C) can be achieved only if the ambient temperature is >20°C.

You can change the temperature set point by pressing the up (▲) or the down (▼) arrow keys above and below the word TEMP until the desired set point is

shown in the display. Typically, the microplate chamber will reach 37.0°C in 15 to 30 minutes.

The microplate chamber temperature is maintained at the set point until you press the incubator

[TEMP On/Off] key again, turning temperature regulation

off. The microplate drawer will open and the temperature within the chamber will begin returning to ambient.

384

has been designed to regulate the temperature

[TEMP On/

384

micro-

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Microplate Spectrophotometer Operator’s Manual 1-11

Chapter 1

NOTE:

Should you turn the incubator back on after a momentary shutdown, allow about ten minutes after reaching temperature for the control algorithm to fully stabilize the microplate chamber temperature.

Temperature regulation and control of the microplate/cuvette chamber are achieved through electric heaters, a fan, efﬁcient insulation, and temperature sensors. The heaters are located in the chamber which is insulated to maintain the temperature set point. The sensors, also mounted inside the chamber, measure the air temperature. The temperature feedback closed-loop control algorithms compare the measured air temperature inside the chamber against the temperature set point, and use the difference to calculate the heating cycles. This technique results in accurate, precise control of the chamber temperature with a temperature variation of the air inside the chamber of less than 1.0°C. (The temperature uniformity within the microplate or cuvette will depend upon the design, materials, and/or conﬁguration of that component.)

Data Collection

The SPECTRAmax PLUS buffer memory.

CAUTION:

Data in the buffer memory is lost when power to the SPECTRAmax PLUS outages. Do not turn the instrument off while important data remains in the buffer memory.

384

stores only the most recent cuvette reading in a

384

is turned off. This applies even to short power

Printed Data Output

An optional printer, designed to sit on top of the instrument, can be purchased for use with the SPECTRAmax PLUS

During stand-alone operation, results are automatically printed as soon as a cuvette has been read. A new cuvette can be loaded into the SPECTRAmax

384

PLUS

while the results from the ﬁrst reading are being printed.

384

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

Figure 1.8: Sample Printout from Thermal Printer

NOTE:

If you have performed a REF reading, the REF values will not be shown as raw OD values, but are used to calculate subsequent sample reads which are shown on the printout and display.

Computer Control

The SPECTRAmax PLUS through which a computer can communicate with and control the instrument.

SOFTmax PRO

Molecular Devices’ SOFTmax PRO software is a highly integrated program that can be used to control and collect data from the SPECTRAmax PLUS SOFTmax PRO is easy to use, yet is powerful and ﬂexible, and makes it possible to access the full capabilities of the SPECTRAmax PLUS

SOFTmax PRO allows you to:

• Read microplates for endpoint, kinetic, or spectral scans

- Use up to six wavelengths for endpoint and kinetic readings

- Perform Absorbance or %Transmittance readings in the 190- to 1000-nm range

384

is equipped with an 8-pin DIN RS-232 serial port

384

- Extend kinetic run times up to 99 hours

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Microplate Spectrophotometer Operator’s Manual 1-13

Chapter 1

- Select your own read intervals for kinetic runs

- Specify the duration for Automix before and between readings

- Read the whole plate or a subset of microplate strips

• Read cuvettes for endpoint, kinetic, or spectral scans

- Use up to six wavelengths for endpoint and kinetic readings

- Perform spectral scans from 190 to 1000 nm

- Extend kinetic run time up to 99 hours

- Select your own read intervals for kinetic runs

- Read a reference sample to be used with a single cuvette or a series of cuvettes

• Use PathCheck to normalize the absorbance readings in each microplate well to a 1-cm pathlength

• Design microplate templates and cuvette templates to simplify data reduction

- Identify groups of wells or cuvettes with labels of your choice

- Identify individual wells or cuvettes with unique names

- Blank the entire plate, groups, and/or individual wells

• Save instrument settings, template formats, and data analysis parameters for both microplates and cuvettes as assay protocol ﬁles and recall them for later use

- Rapid instrument and analysis set up for repeated microplate or cuvette assays

- Uniform analysis for equivalent microplates and cuvettes

• Turn the incubator on or off to control the temperature in the microplate drawer and cuvette port

• Acquire data from the SPECTRAmax PLUS

384

- Save data files for in-depth analysis at a later time

- Save multiple microplates and cuvettes with individual template and data

analysis parameters in one or more experiments in a single data file

- Pre-read microplates

- Analyze kinetic and spectrum data as it is collected

• Use the Automix function to shake the microplate at preset intervals, thereby mixing the contents of each well (highly recommended for ELISAs and other solid-phase, enzyme-mediated reactions)

• Display data on screen

- Raw values, reduced number, or raw values with reduced number

- Raw microplate data in a microplate format

- Ranged data as integers between 0 and 9 in a microplate format

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

- Threshold data as being above, below, or between set limits in a microplate

format

- Gray scale data in seven shades of gray corresponding to high and low limits

in a microplate format

- Kinetic or spectrum plots of all 96 or 384 microplate wells

- Enlarge the display of individual well plots and overlay multiple well plots

- Multiple cuvette data as raw, ranged, threshold, gray scale, kinetic, or spectral

scans

• Perform data analysis using SOFTmax PRO features

- Calculate maximum kinetic rates on non-linear data

- Assign plate, group, or sample blanks

- Perform pathlength correction based on 1-cm cuvette readings

- Customize data analysis for each group in the template

- Create graphs with multiple plots

- Pick from nine curve-fitting routines

- Analyze unknown samples against a standard curve

- Analyze and compare data within a plate, between plates, and between exper-

iments

• Multiple print formats

- Print all or individual sections of the data file

- Define and print a report containing only selected sections

- Customize the order of data file sections

• Export data in tab-delimited ASCII format for use with Excel or other database programs or as JCAMP-DX format for use with spectral analysis software

For a complete description of the features of SOFTmax PRO, refer to the

SOFTmax PRO User’s Manual.

SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual 1-15

Chapter 1

Speciﬁcations Thermal speciﬁcations for microplates used in the SPECTRAmax PLUS

to ﬂat-bottom microplates with isolated wells. All other microplate speciﬁcations apply to standard 96-well polystyrene ﬂat-bottom microplates. Performance speciﬁcations for cuvette readings apply only to aqueous solutions having solute molal concentrations less than 0.4 M. When pathlength compensation is applied to microplate absorbance measurements, agreement with cuvette absorbance measurements for the same solution requires that the solution volume in the microplate well be between 100 µL and 300 µL.

NOTE:

Wavelength range 190–1000 nm

Wavelength selection Monochromator tunable in 1-nm increments

Wavelength bandwidth ≤ 2.0 nm FWHM (full width half maximum)

Wavelength accuracy ± 1.0 nm across wavelength range

Wavelength repeatability ± 0.2 nm across all optical channels

Photometric range -0.3 to 4.000 OD

Photometric resolution 0.001 OD

Technical speciﬁcations are subject to change without notice.

Photometric Performance

384

apply

Photometric accuracy/linearity

(microplate), 0–2.0 OD: 190–1000 nm <± 1.0% and ± 0.006 OD

Photometric accuracy/linearity

(cuvette), 0–2.0 OD: 90–1000 nm <± 1.0% and ± 0.005 OD

Photometric precision

(repeatability), 0–2.0 OD: 190–1000 nm <± 1.0% and ± 0.003 OD

Stray light ≤ 0.05% at 230 nm

Photometric stabilization Instantaneous

Photometric drift None—continuous referencing of monochromatic output

Calibration Automatic before first kinetic read and before every endpoint

reading

Optical alignment None required

Light source Xenon flash lamp (5 watts)

Average lamp lifetime 1 billion flashes

Illumination Top down (microplates); horizontal (cuvettes)

Photodetectors Silicon photodiode

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

Photometric Analysis Modes

Stand-Alone

• Single wavelength Absorbance or %Transmittance reading of the cuvette (or test tube)

Using SOFTmax PRO

• Express data as Absorbance or %Transmittance

• Single wavelength reading of microplate and/or cuvette

• Multiple wavelength (up to six) reading of microplate and/ or cuvette

• Kinetic and kinetic graphics of microplate and/or cuvette

• Spectral scan (190–1000 nm) of microplate and/or cuvette

Measurement Time (calibration off)

Microplate

Read time (endpoint)

-- Standard read • 96 wells in 9 seconds (single wavelength)

• 96 wells in 9*N seconds (N wavelengths)

• 384 wells in 29 seconds (single wavelength)

• 384 wells in 29*N seconds (N wavelengths)

-- Speed read • 96 wells in 5 seconds (single wavelength)

• 96 wells in 5*N seconds (N wavelengths)

• 384 wells in 16 seconds (single wavelength)

• 384 wells in 16*N seconds (N wavelengths)

Kinetic read intervals • 96 wells, 9-second minimum interval between readings

(single wavelength)

• 1 column, 2-second minimum interval between readings (single wavelength)

• 384 wells, 29-second minimum interval between readings (single wavelength)

Cuvette

Read time (endpoint) • 1 second (single wavelength)

Kinetic read intervals • 2-second minimum interval between readings

(single wavelength)

Cuvette: Normal scan 45*K nm/min (K = wavelength interval)

Cuvette: Speed scan 130*K nm/min

Microplate: Normal scan 33*K nm/min (8-well strip); 21*K nm/min (16-well strip)

Microplate: Speed scan 135*K nm/min (8-well strip); 77*K nm/min (16-well strip)

SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual 1-17

Scan Speed

, 0 –

Chapter 1

Speciﬁcations

Wavelength range 190–1000 nm

Wavelength selection Monochromator tunable in 1-nm increments

Wavelength bandwidth

Wavelength accuracy ± 1.0 nm across wavelength range

Wavelength repeatability ± 0.2 nm across all optical channels

Photometric range -0.3 to 4.000 OD

Thermal speciﬁcations for microplates used in the SPECTRAmax PLUS to ﬂat-bottom microplates with isolated wells. All other microplate speciﬁcations apply to standard 96-well polystyrene ﬂat-bottom microplates. Performance speciﬁcations for cuvette readings apply only to aqueous solutions having solute molal concentrations less than 0.4 M. When pathlength compensation is applied to microplate absorbance measurements, agreement with cuvette absorbance measurements for the same solution requires that the solution volume in the microplate well be between 100 µL and 300 µL.

NOTE:

Technical speciﬁcations are subject to change without notice.

384

apply

Photometric Performance

≤

2.0 nm FWHM (full width half maximum)

Photometric resolution 0.001 OD

Photometric accuracy/linearity

(microplate)

Photometric accuracy/linearity

(cuvette)

Photometric precision

(repeatability)

Photometric stabilization Instantaneous

Photometric drift None—continuous referencing of monochromatic output

Optical alignment None required

Average lamp lifetime 1 billion flashes

Photodetectors Silicon photodiode

2.0 OD: 190–1000 nm <± 1.0% and ± 0.006 OD

2.0 OD: 90–1000 nm <± 1.0% and ± 0.005 OD

2.0 OD: 190–1000 nm <± 1.0% and ± 0.003 OD

Stray light

Calibration Automatic before first kinetic read and before every endpoint reading

Light source Xenon flash lamp (5 watts)

Illumination Top down (microplates); horizontal (cuvettes)

≤

0.05% at 230 nm

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Microplate Spectrophotometer Operator’s Manual

Instrument Description

Photometric Analysis Modes

Stand-Alone

• Single wavelength Absorbance or %Transmittance reading of the cuvette (or test tube)

Using SOFTmax PRO

• Express data as Absorbance or %Transmittance

• Single wavelength reading of microplate and/or cuvette

Multiple wavelength (up to six) reading of microplate and/or cuvette

•

• Kinetic and kinetic graphics of microplate and/or cuvette

• Spectral scan (190–1000 nm) of microplate and/or cuvette

Measurement Time (calibration off)

Microplate

Read time (endpoint)

-- Standard read* • 96 wells in 9 seconds (single wavelength)

• 96 wells in 19 seconds (dual wavelength—425 & 650 nm)

• 384 wells in 29 seconds (single wavelength)

• 384 wells in 59 seconds (dual wavelength—425 & 650 nm)

Speed read* • 96 wells in 5 seconds (single wavelength)

• 96 wells in 12 seconds (dual wavelength—425 & 650 nm)

• 384 wells in 16 seconds (single wavelength)

• 384 wells in 34 seconds (dual wavelength—425 & 650 nm)

Kinetic read intervals • 96 wells, 9-second minimum interval between readings

(single wavelength)

• 1 column, 2-second minimum interval between readings (single wavelength)

• 384 wells, 29-second minimum interval between readings (single wavelength)

Cuvette

Read time (endpoint) • 1 second (single wavelength)

Kinetic read intervals • 2-second minimum interval between readings

(single wavelength)

*Measurement conditions: endpoint, column priority (for dual-wavelength measurements), calibrate off.

Cuvette: Normal scan 45*K nm/min (K = wavelength interval)

Cuvette: Speed scan 130*K nm/min

Microplate: Normal scan 33*K nm/min (8-well strip); 21*K nm/min (16-well strip)

Microplate: Speed scan 135*K nm/min (8-well strip); 77*K nm/min (16-well strip)

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Microplate Spectrophotometer Operator’s Manual 1-17

Scan Speed

Chapter 1

Temperature Regulation

Reading chamber Isothermal when temperature regulation is not enabled

Range 4°C above ambient to 45°C when temperature regulation enabled.

The ambient temperature must be >20°C to achieve temperature regulation at 45°C.

Resolution ± 0.1°C

Accuracy ± 1.0°C for microplate and cuvette chamber

Temperature uniformity at equilibrium ± 0.5°C at 37°C

Chamber warm-up time 15-30 minutes (measured on air) after initiation of temperature

regulation

Temperature regulation 4 sensors

Drift ± 0.2°C (regulated)

Temperature regulation diagnostics

Evaporation Plate lid required to minimize evaporative cooling

Recommended microplate Flat-bottom microplates with isolated wells and lid

Temperature regulation system is continuously monitored and updated

Automix with SOFTmax PRO

Plate mixing modes Selectable: off, once prior to any reading, and once prior to and be-

tween kinetic readings

Plate mixing duration Selectable: 1 to 999 seconds (three-second default)

Compatibility

Microplate Standard and half-area 96-well flat-bottomed microplates (0.3 mL).

384-well flat-bottomed microplates. Polystyrene plates for wavelengths above 340 nm; UV transparent plates above 220 nm; quartz plates above 190 nm.

Cuvette Standard height (45 mm) cells with 10 mm pathlength (12.5 mm x

12.5 mm outside) with minimum inside width of 4 mm (typical for 3 mL volume cells).

Test Tubes 12 x 75 mm test tubes can be used in the cuvette chamber with the

test tube cover.

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Microplate Spectrophotometer Operator’s Manual

General Instrument

Instrument Description

Display 2-

Operating panel 9-key membrane keypad

Self-diagnosis Continuous on-board diagnostics

Spill control Drawer mechanism/reading chamber assembly is protected from ac-

Computer interface 8-pin DIN RS-232 serial (double shielding required)

Printer interface Parallel 25-pin to Centronics (double shielding required)

Microplates supported All 96-well and strip-well microplates, including lids

-20-character backlit LCD

cidental spillage by drainage ports

Environmental

Operating temperature 15 to 40°C

Operating humidity 0 to 70%, non-condensing

Storage temperature -20 to 65°C

Physical

Size (h

8.6 in. (22 cm)

Weight 30 lb (13.6 kg)

22.8 in. (58 cm)

15 in. (38 cm)

Power consumption < 250 watts

Line voltage 90-250 VAC, auto-ranging

Line frequency 50/60 Hz

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Microplate Spectrophotometer Operator’s Manual 1-19

Chapter 2

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Microplate Spectrophotometer Operator’s Manual

Installation

Installation Warnings

Installation Cautions

1) Always make sure the power switch on the instrument is in the OFF position and remove the power cord from the back of the instrument prior to any installation or relocation of the instrument.

2) Do not operate the instrument in an environment where potentially damaging liquids or gases are present.

1) Do not touch or loosen any screws or parts other than those speciﬁcally designated in the instructions. Doing so might cause misalignment and will void the instrument warranty.

Unpacking The SPECTRAmax PLUS

retain the carton and the packing materials. If the unit should need to be returned for repair, you must use the original packing materials and carton for shipping. If the carton has been damaged in transit, it is particularly important that you retain it for inspection by the carrier in case there has also been damage to

the instrument.

WARNING:

(13.6 kg) and should be lifted with care. It is recommended that two persons lift the instrument together, taking the proper precautions to avoid injury.

The SPECTRAmax PLUS

384

is packed in a specially designed carton. Please

384

weighs approximately 30 lb

After examining the carton, place it on a ﬂat surface in the upright position. Open the top of the box and lift the SPECTRAmax PLUS packing materials around the ends, up and out of the shipping box. Remove the packing material from both ends of the instrument and set the instrument down carefully.

384

, along with the

SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual 2-3

Chapter 2

Setting Up for StandAlone Use

Fuse Box Cover

Label

1) Place the SPECTRAmax PLUS

384

on a level surface, away from direct sunlight,

dust, drafts, vibration, and moisture.

2) Turn the instrument around so that the back of the instrument is facing you as shown in Figure 2.1.

Power Switch

Power Cord

Receptacle

Printer (Centronics)

Computer (RS-232)

Figure 2.1: View of Rear Panel

3) Insert the appropriate (Macintosh or PC) serial interface cable into the RS232 serial port on the SPECTRAmax PLUS

384

. Connect the other end to the

serial port of the computer.

4) Insert the female end of the power cord into the power receptacle at the rear of the SPECTRAmax PLUS

384

. Connect the male end to a grounded power outlet of the appropriate voltage. Molecular Devices recommends that you use a surge protector between the power cord and the grounded power outlet.

5) If you are using a printer for reading cuvettes in stand-alone mode, locate the printer port (25-pin parallel) on the rear panel of the SPECTRAmax

384

PLUS

. Connect one end of the cable to this port and the connect the other (Centronics) end to the printer. Load paper into the printer according to the manufacturer’s instructions and connect the printer’s power cord to the power outlet.

6) Turn the SPECTRAmax PLUS

384

around so that the control panel now faces you. Be sure no cables run beneath the instrument. Leave at least three inches between the back of the instrument and the nearest objects or surfaces to ensure proper ventilation and cooling.

7) Remove the tape from the cuvette door.

384

8) Turn on the power to the SPECTRAmax PLUS

, wait for the microplate drawer to open, and remove the tape and protective covering from the drawer subplate.

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Chapter 3 Stand-Alone Operation

Prepare for a Reading . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

Turn the Instrument and Printer On . . . . . . . . . . 3-3

Set the Temperature. . . . . . . . . . . . . . . . . . . . . . . 3-3

Select the Wavelength . . . . . . . . . . . . . . . . . . . . . 3-3

Read the Cuvette . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Operation Overview. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-4

Chapter 3

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Microplate Spectrophotometer Operator’s Manual

Stand-Alone Operation

Prepare for a Reading

This chapter contains operating information for the SPECTRAmax PLUS plate/Cuvette Spectrophotometer. If you are an experienced user of this instrument, you can turn to the Operation Overview on page 3-4 for a quick review of the operating steps.

NOTE:

Only temperature selection and single-wavelength cuvette readings are

384

Micro-

available in stand-alone mode through front panel control. To achieve full use of this instrument, connect it to a computer and use SOFTmax PRO software.

Turn the Instrument and Printer On

The power switch for the SPECTRAmax PLUS Press the rocker switch to the ON position. The instrument will automatically perform diagnostic checks to ensure that it is functioning correctly. Turn the printer on at the same time.

384

is located on the back panel.

Set the Temperature

If elevated temperature within the cuvette or microplate chamber is required, you should turn on the incubator ﬁrst, allowing enough time for the temperature to reach the set point before performing a reading. When you ﬁrst turn the instrument on, up to 30 minutes may be required for the temperature within the chamber to reach the set point.

To enable the incubator, press the incubator

[TEMP On/Off] key. The LCD dis-

play will update to show the temperature set point and the current chamber temperature.

To change the temperature set point, press the up or down arrows above and below TEMP until the desired temperature set point is shown in the display.

The microplate/cuvette chamber temperature will be maintained at the set point until you disable temperature control by touching the incubator

[TEMP On/Off] key again. When the incubator is off, the drawer will open

and the temperature within the chamber will begin returning to ambient.

NOTE:

Should you turn the incubator back on after a momentary shutdown, allow about ten minutes for the control algorithm to fully stabilize the chamber temperature.

Select the Wavelength

Press the up or down arrow keys above and below the λ to increment or decre-

ment the wavelength setting (1 nm increments).

SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual 3-3

Chapter 3

Read the Cuvette

Operation Overview

After setting the temperature (if desired), and choosing the wavelength for the reading, insert the cuvette into the chamber, making sure that the clear sides are to the left and right (facing the instrument). Do not touch the clear surfaces of the cuvette. Make sure the cuvette is completely seated in the chamber and close the cuvette door. If the cuvette contains a blank (typically this solvent contains everything that the samples contain except for analyte), press the acquire the reference reading from the cuvette. The SPECTRAmax PLUS

[REF] key to

384

will automatically calibrate in less than two seconds, send a header for the reading to the printer, close the microplate drawer (if it was open), and read the cuvette according to the selected instrument settings. If the cuvette contains a sample, touch the

[READ CUVETTE] key to acquire the sample reading from the

cuvette. When reading is complete, remove the cuvette.

The following steps provide a quick reminder of the basic operating procedures required to read a cuvette using the SPECTRAmax PLUS

1) Turn on the power switch of the SPECTRAmax PLUS

384

(located on the back

panel).

2) If you wish to regulate the temperature inside the microplate reading chamber, touch the

[TEMP On/Off] key to bring the chamber to the default temperature

of 37.0°C. The microplate drawer will close and the indicator on the right of the LCD will ﬂash until the set temperature is reached.

3) If the incubator is on, the LCD will show the current temperature along with the temperature set point. To change the set point (to any setting from ambient +4° to 45°C), press either the up or down arrow above or below TEMP.

4) Select the desired measurement wavelength by pressing the up or down arrow

near λ. Scroll up or down through the list of wavelengths shown in the LCD

using the up or down arrows until the desired measurement wavelength is highlighted.

5) Load the prepared cuvette into the chamber, being sure that the clear sides are left and right (when facing the instrument). Press the

[REF] or [READ CUVETTE]

key.

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Microplate Spectrophotometer Operator’s Manual

Chapter 4 Maintenance and Troubleshooting

Technical Support . . . . . . . . . . . . . . . . . . . . . . . . 4-3

Warnings and Cautions . . . . . . . . . . . . . . . . . . . . 4-3

General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4

Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4

Changing the Fan Filter . . . . . . . . . . . . . . . . . . . . 4-4

Changing the Fuses . . . . . . . . . . . . . . . . . . . . . . . 4-5

Error Codes and Probable Causes. . . . . . . . . . . . . 4-7

Error Messages . . . . . . . . . . . . . . . . . . . . . . 4-7

Opening the Drawer Manually . . . . . . . . . . . . . 4-11

Chapter 4

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Microplate Spectrophotometer Operator’s Manual

Maintenance and Troubleshooting

This chapter lists maintenance procedures and troubleshooting information, including error codes that may be seen, followed by their most likely causes and remedies.

Technical Support

Warnings and Cautions

Molecular Devices Corporation is a leading worldwide manufacturer and distributor of analytical instrumentation. We are committed to the quality of our products and to fully supporting our customers with the highest level of technical service. In order to fully beneﬁt from our technical services, please complete the registration card and return it to the address printed on the card.

If you have any problems using the SPECTRAmax PLUS

384

Microplate Spectrophotometer that are not covered in this chapter, in the U.S., contact our Technical Services group at 1-800-635-5577; elsewhere contact your local representative.

BIOHAZARD:

It is your responsibility to decontaminate the instrument, as well as any accessories, before requesting service by Molecular Devices representatives and before returning the instrument or any components to Molecular Devices Corporation.

WARNING:

All maintenance procedures described in this manual can be safely performed by qualiﬁed personnel. Maintenance not covered in this manual should be performed by a Molecular Devices representative.

WARNING:

Removal of protective covers that are marked with the High Voltage warning symbol shown below can result in a safety hazard.

SPECTRAmax PLUS

WARNING:

Always turn the instrument off and remove the power cord and any computer/printer cables from the back prior to any maintenance or installation operation

WARNING:

Never perform any operation on the instrument in an environment where liquids or potentially damaging gases are present.

WARNING:

CAUTION:

harm to the optics in the SPECTRAmax PLUS

Risk of electrical shock. Refer servicing to qualiﬁed personnel.

Use of organic solvents (such as dichloromethane) may cause

384

. Extreme caution is advised when using organic solvents. Always use a plate lid and avoid placing a plate containing these materials in the reading chamber for prolonged periods of time. Damage caused by the use of incompatible or aggressive solvents is NOT covered by the instrument warranty.

CAUTION:

Never touch any of the optic mirrors, ﬁlters, or cables or their housing, or manifold. The optics are extremely delicate, and critical toe of the instrument.

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Do not touch or loosen any screws or parts other than those spe-

CAUTION:

ciﬁcally designated in the instructions. Doing so could cause misalignment and possibly void warranty.

General Keep the drawer closed when the instrument is not in use. The drawer can be

opened by pressing the DRAWER button. Always close the drawer immediately prior to switching the instrument off.

Cleaning Wear gloves during any cleaning procedure that could

Cleaning the Fan Filter

BIOHAZARD:

involve contact with either hazardous or biohazardous materials or ﬂuids.

Periodically, you should clean the outside surfaces of the SPECTRAmax PLUS using a cloth or sponge that has been dampened with water. Do not use abrasive cleaners. If required, clean the surfaces using a mild soap solution diluted with water or a glass cleaner and then wipe with a damp cloth or sponge to remove any residue. Do not spray cleaner onto the instrument.

If needed, clean the microplate drawer or cuvette chamber using a cloth or sponge that has been dampened with water.

Should ﬂuids spill in the drawer area (when the drawer is out) or into the cuvette chamber (when the door is open), they will be directed to a tray at the bottom of the instrument from which they will exit to the bench or counter beneath the instrument. Wipe up any spills immediately. Clean only the exterior of the unit (and the microplate drawer or inside the cuvette chamber if necessary). Never clean the inside of the instrument. Do not allow excess water or other ﬂuids to drip inside the instrument.

The fan ﬁlter on the bottom of the instrument requires periodic cleaning. The frequency of the cleaning depends on how dusty your particular lab is and could range from once a month to once every six months.

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1) Turn power to the instrument OFF and then remove the power cord and cables from the back of the instrument.

2) Make sure no plate or cuvette is in the instrument. Tape the cuvette door closed. Turn the instrument over so that it rests ﬂat on the bench.

3) Pop the black fan cover off the remove the ﬁlter.

4) Clean the ﬁlter by blowing clean, canned air through it or by rinsing it—ﬁrst with water and then with alcohol—and allowing it to dry completely.

5) Place the clean, dry ﬁlter over the fan and replace the black cover.

6) Turn the instrument back over. Remove the tape from the cuvette door. Reconnect the power cord and cables to the instrument.

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Maintenance and Troubleshooting

Changing the Fuses

Fuses burn out occasionally and must be replaced. If the instrument does not seem to be getting power after switching it on (the LCD shows no display), ﬁrst check to see whether the power cord is securely plugged in to a functioning power outlet and to the receptacle at the rear of the SPECTRAmax PLUS failed while the SPECTRAmax PLUS

384

was already on, check that the power

384

. If power

cord is not loose or disconnected and that power to the power outlet is functioning properly. If these checks fail to remedy the loss of power, follow the steps listed below to replace the fuses. Spare fuses (two U.S. and two metric) are shipped with the instrument. The U.S. and metric fuses are identical except for physical size. They may be taped to the back of the SPECTRAmax PLUS

384

If you no longer have spare fuses, you may obtain new ones from Molecular Devices (part numbers: 4601-0013 for U.S., 4601-0014 for metric) or from a local hardware store. Make sure fuses are rated SLOWBLOW (U.S.: 4-amp time-

delay; metric: 4-amp, 5 × 20 mm, time-delay).

To change fuses, follow the steps below.

1) Switch power to the instrument off and then remove the power cord from the outlet and from the SPECTRAmax PLUS

384

power cord receptacle.

2) Remove the printer cable and computer cable (if connected) from the back of the SPECTRAmax PLUS

384

3) Turn the instrument around for easy access to the rear panel.

4) On the left-hand side of the rear panel (viewed from the back) is the power switch, fuse box, and power cord receptacle. As shown in the ﬁgures below, press to the right of the black plastic cover of the fuse box to release it. Pull the fuse box cover away from the instrument. The fuse box will begin to slide forward.

Fuse Box Cover

Figure 4.1: Power Switch, Fuse Box, and Power Receptacle

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Chapter 4

5) Continue gently pulling the fuse box forward until it is free of the instrument.

Figure 4.2: Removing the Fuse Box

6) When removed, the fuse assembly will appear as shown in Figure 4.3. The holder inside contains two fuses.

7) It is possible that only one of the fuses may have blown. Molecular Devices recommends that you replace both fuses, however, to ensure continued proper operation. Pull both fuses out of the holder and discard them.

Figure 4.3: The Fuse Box and Holder (with Fuses) Removed

8) Insert new SLOWBLOW-rated fuses into the fuse holder. Either end of the fuse may be forward.

9) Insert the fuse box into the opening in the instrument, making sure that the fuses are on the right side (toward the power receptacle). Press the fuse box into place, making sure the cover snaps closed.

10) Reconnect the power cord to the instrument and to the wall outlet and reconnect other cables previously disconnected.

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Maintenance and Troubleshooting

Error Codes and Probable Causes

If a problem occurs during operation that causes an unrecoverable error, the instrument will stop and an error code number will be shown in the display on the front panel. To correct the problem, call your local Molecular Devices representative for assistance.

Error Messages

The LCD will display Fatal Error codes when a situation arises that requires attention. Any reading in progress will stop. Warning messages do not stop a read- ing but are logged in the error buffer. Warning messages indicate a situation that requires attention but is not sufﬁcient to stop or prevent a reading. Examples of situations that might cause warning messages are low memory, entries being out of range, or operations that could result in loss of data. These messages are generally self explanatory. For assistance regarding warning messages, contact your local Molecular Devices representative.

Error Code Classiﬁcations

Not all error messages are listed in this manual. The errors are grouped in relationship to possible causes as follows:

Error Code Numbers Possible Causes

100 -199 Errors possibly caused by unrecognized commands being sent from the computer

to the instrument.

200-299 Errors probably due to a main board failure or an error in the ﬁrmware code.

Most of these errors require the assistance of Technical Support.

300-399 Instrument errors due to either a main board failure or other system failure. Most

of these errors require the assistance of Technical Support.

400-499 Errors caused by a motor motion failure. Most of these errors require the assis-

tance of Technical Support.

500-599 Errors due to failure or improper initialization of the instruments non-volatile

memory (NVRAM). All of these errors require the assistance of Technical Support.

Some errors (shown in boldface in this manual) are considered fatal if they are detected during power up (the instrument will abort the power up sequence and display “FATAL ERROR ###” on the LCD panel). Check the following table to see if you can do something to prevent the fatal error (e.g., closing the cuvette door during the power-up sequence). After correcting the problem, leave the instrument on for about ﬁve minutes, turn it off and then back on. If you continue to get the fatal error message on power up, record the error message number and contact Molecular Devices Technical Support or your local representative for assistance.

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Chapter 4

NOTE:

If the instrument is functioning normally when using SOFTmax PRO,

no errors should be in the buffer (except error number 100).

Table 4.1. Error Codes, Messages, and Notes about the Errors

ERROR

CODE

100-199: Operator Errors

100 command not found Command string not recognized.

101 invalid argument Command Argument not recognized.

102 too many arguments Too many arguments after command.

103 not enough arguments Missing arguments.

104 input line too long Too many characters in the input line.

105 command invalid, sys-

ERROR MESSAGE NOTES

Instrument could not perform the give command because it was

tem busy

busy doing another task. Example: Request a wavelength while the monochromator is in motion.

106 command invalid, mea-

surement in progress

Instrument could not perform command because a measurement was in progress

107 no data to transfer Inputting transfer when there’s no data in the buffer

108 data buffer full To many data sets in the buffer. Can be caused by setting up a

long kinetic and disconnecting computer or SOFTmax PRO is preempted by another application.

109 error buffer overﬂow More than 65 errors in the buffer, clear the buffer.

110 stray light cuvette, door

Cuvette door open while doing a read.

open?

111 invalid read settings

200-299: Firmware Errors

200 assert failed Firmware error.

201 bad error number Firmware error.

202 receive queue overﬂow Caused by external device sending too much data over serial port

and ignoring ﬂow control.

203 serial port parity error Parity bit error detected with incoming serial data.

204 serial port overrun error Caused by host computer sending too much data and ignoring

205 serial port framing error

4-8 SPECTRAmax PLUS

the ﬂow control signal.

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Table 4.1. Error Codes, Messages, and Notes about the Errors

Maintenance and Troubleshooting

ERROR

CODE

206 cmd generated too

ERROR MESSAGE NOTES

Firmware error.

much output

207 fatal trap Instrument error. Instrument locks up.

208 RTOS error Firmware error.

209 stack overﬂow Firmware error.

210 unknown interrupt Firmware error.

300-399: Hardware Errors

300 thermistor faulty Unable to read a reasonable thermistor value. Thermistor faulty

or disconnected, Main board problem, or ambient temperature out of range.

301 safe temperature limit

exceeded

A temperature of over 50°C detected on one or more of the 4 thermistors. Temperature will be shut off and remain off until a successful completion of power-up reset.

302 low light Not enough light detected to make an accurate measurement. If

doing a cuvette read, the cuvette door may be open.

303 unable to cal dark cur-

rent

Too much stray light detected on power-up, faulty or disconnected pre-amp boards.

304 signal level saturation During a cuvette read, could be due to cuvette door being open.

305 reference level saturation During a cuvette read, could be due to cuvette door being open.

306 plate air cal fail, low

Minimum signal/reference ratio not met during air calibration.

light

307 cuv air ref fail

308 stray light Light leak in reading chamber or cuvette door open. Could also

be a faulty pre-amp board.

309 front panel not respond-

LCD front panel bad or disconnected.

ing

310

311

312 gain calibration failed Power-up calibration and check of signal path gain is out of tol-

erance. Could be due to bad or disconnected pre-amp or excessive stray light.

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Chapter 4

Table 4.1. Error Codes, Messages, and Notes about the Errors

ERROR

CODE

ERROR MESSAGE NOTES

313 reference gain check fail Power-up check of the Reference ampliﬁer's gain out of toler-

ance. Could be due to bad or disconnected pre-amp board or excessive stray light.

314 low lamp level warning

315 can't ﬁnd zero order On power-up grating motor could not ﬁnd zero-order home

position.

316 grating motor driver

faulty

Grating motor didn’t move to where it was commanded to in a reasonable time.

317 monitor ADC faulty

400-499: Motion Errors

400 carriage motion error Carriage did not move to either of its photo interrupts in a rea-

sonable time, or can’t ﬁnd its photo interrupt.

401 ﬁlter wheel error Filter wheel did not move to its photo interrupt in a reasonable

time, or can’t ﬁnd its photo interrupt.

402 grating error Grating did not move to its photo interrupt in a reasonable

time, or can’t ﬁnd its photo interrupt.

403 stage error Stage did not move to its photo interrupt in a reasonable time,

or can’t ﬁnd its photo interrupt.

500-599: NVRAM Errors

500 NVRAM CRC corrupt The CRC for the NVRAM data is corrupt.

501 NVRAM Grating cal

Grating calibration data is unreasonable.

data bad

502 NVRAM Cuvette air cal

Cuvette air calibration data is unreasonable.

data error

503 NVRAM Plate air cal

Plate air calibration data is unreasonable.

data error

504 NVRAM Carriage offset

Carriage offset data is unreasonable.

error

505 NVRAM Stage offset

Stage offset data is unreasonable.

error

506 NVRAM Battery Time to replace the NVRAM battery (U3).

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Microplate Spectrophotometer Operator’s Manual

Maintenance and Troubleshooting

Opening the Drawer Manually

If an error occurs while the drawer is closed and you need to open the drawer, press the ment off and then on again.

If the drawer still remains closed, turn the power off and using your thumb nail, locate the groove in the upper left side wall of the door. Open the door, and with your index ﬁnger, pull the microplate drawer out of the instrument (do not force the drawer) and remove the microplate. This action will not harm the instrument, but should only be taken if the ﬁrst two options have failed to open the drawer.

If you are still unable to open the drawer, contact Molecular Devices Technical Support or your local representative for assistance.

[Drawer] key. If the drawer does not open, turn power to the instru-

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Chapter 4

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Maintenance and Troubleshooting

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Chapter 4

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Maintenance and Troubleshooting

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Chapter 4

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Maintenance and Troubleshooting

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Chapter 4

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Microplate Spectrophotometer Operator’s Manual

Appendix A Printers and Cables

Compatible Printers . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3

Cables . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . A-3

Appendix A

A-2 SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Printers and Cables

Compatible Printers

The SPECTRAmax PLUS printer. Compatibility may require changing the dip switch or pin settings on the

384

can be used with an Epson-compatible strip

printer or insertion of an emulation cartridge. Please check your printer’s manual to assess compatibility based on this criterion—some printers will not meet this requirement.

Please contact your local Molecular Devices representative for information regarding recommended printers.

Cables Molecular Devices recommends that you use high-quality, double-shielded cables

to connect the SPECTRAmax PLUS printer or computer. Choose cables that meet the following requirements:

Printer Cable (for Stand-alone operation only):

Centronics parallel, male 25D to Centronics.

Serial Interface Cable

(contact Molecular Devices for speciﬁc pin-out requirements)

Macintosh: Male DB8 to male DB8 IBM Compatible: Male DB8 to Female DB9 (custom cable made by

Molecular Devices)

384

to other peripheral instruments, such as a

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Microplate Spectrophotometer Operator’s Manual A-3

Appendix A

A-4 SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Appendix B Accessories

Items Available for Use with the SPECTRAmax PLUS

384

. . . . . . . . . . . . . . . . . . . . . . . . B-3

Appendix B

B-2 SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Items Available for Use with the

SPECTRAmax

PLUS

384

Accessories

Part Number

Cable, RS-232, Macintosh to SPECTRAmax PLUS

8-pin DIN to 8-pin DIN. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0091

Cable, RS-232, PC to SPECTRAmax PLUS

9-pin to 8-pin DIN . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0149

Cable, parallel printer, 25-pin to Centronics . . . . . . . . . . . . . . . . . . . 9000-0003

Cuvette Calibration Kit . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0161

Fuse, 4-amp Time Delay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4601-0013

Fuse, 4-amp (5 × 20 mm) Time Delay. . . . . . . . . . . . . . . . . . . . . . . . 4601-0014

384

Power Cord (US, Canada, Japan, Mexico, India) . . . . . . . . . . . . . . . . 4400-0002

Power Cord (Germany, France, Scandinavia, Italy, Korea) . . . . . . . . . 4400-0036

Power Cord (UK, Indonesia, Singapore, Malaysia). . . . . . . . . . . . . . . 4400-0037

Power Cord (Australia, Hong Kong, China) . . . . . . . . . . . . . . . . . . . 4400-0038

Printer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0189

SPECTRAplate—Disposable

UV-transparent disposable microplates (case of 50) . . . . . . . . . . . . . . R9013

SPECTRAplate—Quartz

UV-transparent quartz microplate . . . . . . . . . . . . . . . . . . . . . . . . . . . R8024

SPECTRAmax Mouse Pad. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9000-0133

384

SPECTRAtest PLUS

Test tube cover. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2300-0277

Validation Package . . . . . . . . . . . . . . . . . . . . 0200-2405

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Appendix B

B-4 SPECTRAmax PLUS

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Microplate Spectrophotometer Operator’s Manual

Index

Appendix B

Compatible Printers A-1

Appendix C

Accessories B-1

Back Panel 1-10 Bandwidth 1-16

Cables 2-4, A-3

connecting 2-4 serial 1-10

Calibration 1-16 Chapter 1

Instrument Description 1-1

Chapter 3

Stand-Alone Operation 3-1

Chapter 4

Maintenance 4-1 Cleaning 4-4 Component Description 1-5 Computer Control 1-13 Computer control 1-13 Connections 1-5, 1-10

cables 2-4, A-3

printer 1-10, 2-4 Control panel 1-5 Control panel display 1-7 Cuvette 1-9 Cuvette chamber 1-5, 1-9

Error Messages 4-7 Error messages 4-7

Fan

filter cleaning 4-4 Fan filter 4-4 Functional Description 1-11 Fuse 4-5

changing 4-5 Fuses

accessing 1-10

If 3-4 Incubator 1-8, 1-11, 3-3 Installation 2-3 Installation Cautions/Warnings 2-3

Key

READ 3-4 Keys 1-6 Keys (control panel) 1-5

Labels 1-10 LCD (Liquid Crystal Display) 1-6 LCD (liquid crystal display) 4-7 LCD (liquid Crystal Display} 3-4 LCD Display 1-6

Data

collection 1-12 printed output 1-12

Data Collection 1-12 Drawer

open manually 4-11

Drawer key 1-7, 1-8, 1-11

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Microplate Spectrophotometer Operator’s Manual I-1

Microplate 1-3

96-well 1-8

filter-bottom 1-8

strip wells 1-8

temperature uniformity 1-8 Microplate drawer 1-5, 1-7, 1-8

closing 1-8

incubator status 1-8 Microplates 1-8

Index

Opening the Drawer Manually 4-11 Operation Overview 3-4

Pathlength correction 1-4 Ports 2-4

printer 1-10

serial 1-10 Power cord 1-10, 2-4, B-3 Power switch 1-5, 1-10, 3-3 Preparing for a Reading 3-3 Print 1-12 Printed Data Output 1-12 Printer 1-4, 1-12, 1-13, 2-4, 3-3 Printer port 1-10, 2-4

Read mode 1-13, 3-4 Read the Microplate 3-4 Read the microplate 1-13, 1-14 Reference reading 1-7, 3-4 Remote control 1-13

Technical service 4-3 Technical Support 4-3 Temperature 1-11, 1-12, 3-4

equilibration 1-12, 3-3 incubator 1-12, 3-3 regulation 1-11 set point 1-6, 1-12, 3-3 setting 3-3

stabilization 1-12 Temperature Regulation 1-11 The Back Panel 1-10 The Microplate Drawer 1-8, 1-9 Turn the Instrument On 3-3

Unpacking 2-3

Warranty ii Wavelength 3-4

range 1-16

Selection 1-11 Wavelength Selection 1-11 Wavelength setting 1-6

Sample (cuvette) reading 1-7 Select the Wavelength(s) 3-3 Serial port 1-10 Set point 1-6 Set point, temperature 1-7 Set the Temperature 3-3 Setting Up for Stand-Alone Use 2-4 SOFTmax PRO 1-3, 1-5, 1-13 Software

SOFTmax PRO 1-13 Specifications 1-16 Spills 4-4

cleaning 4-4 Stand-alone operation 1-11, 2-4

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Molecular Devices SPECTRAmax PLUS 384 Operator's Manual

Specifications and Main Features

Frequently Asked Questions

User Manual