Molecular Devices SpectraMax 190 User manual

SPECTRAmax
®
190
Microplate Spectrophotometer
Operator’s Manual
Molecular Devices Corporation
1311 Orleans Drive
Sunnyvale, California 94089
Part # 0112-0085
Rev. A

ii SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
Molecular Devices Corporation
SPECTRAmax
®
190 Operator’s Manual
Copyright
 Copyright 2001, 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, chem-
ical, 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 190 and methods are protected by the following patents:
U.S. Patents
4,968,148, 5,112,134, 6,151,111, and 6,188,476 ; Canadian Patent 1,305,334 .
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 190 Microplate Spectrophotometer are warranted for a period of
one (1) 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 190 instrument only, and does not extend to any com-
puter, printer, software, reagents, or disposables used with the instrument.

SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual iii
Labor and Parts
To obtain warranty service during the applicable warranty period, you must take the product or de-
liver 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 Devic-
es Corporation service facility to schedule warranty service. You may call Molecular Devices Corpo-
ration at the telephone number or address below to locate the nearest service facility. You must
schedule warranty service
prior to bringing or shipping the product for servicing. At the time of re-
questing 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.
This warranty covers only defects arising under normal usage and does not cover malfunctions or fail-
ures resulting from misuse, abuse, neglect, alteration, modification, or repairs by other than an autho-
rized 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 con-
sequential damages for breach of any express or implied warranty on this product, except to the ex-
tent 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 cov-
ering 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.
If the Buyer furnishes specifications to the Seller, the Buyer agrees to hold the Seller harmless against
any 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 con-
strued 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:
[phone]
[phone] (U.S. and California)
Microplate Spectrophotometer
100-240V ~ 4A 50-60 HZ T4.0A
!
Sunnyvale, CA!
Made in USA
2 Lines Fused, unplug before servicing!
Vor Wartungsarbeiten Netzstecker ziehen!
190
Protected by US Patents 4,968,148; 5,112,134;
6,151,111; and 6,188,476
Protected by Canadian Patent 1,305,334
Other US and International patents pending

iv SPECTRAmax 190 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 . . . . . . . . . . . . . . . . . . . . . 1-7
The Back Panel. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-8
Functional Description. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-9
Stand-Alone Operation . . . . . . . . . . . . . . . . . . . . . 1-9
Wavelength Selection . . . . . . . . . . . . . . . . . . . . . . . 1-9
Temperature Regulation . . . . . . . . . . . . . . . . . . . . 1-9
Computer Control. . . . . . . . . . . . . . . . . . . . . . . . . 1-10
Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-12
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
Turn the Instrument and Printer On . . . . . . . . . . 3-3
Set the Temperature . . . . . . . . . . . . . . . . . . . . . . . . 3-3
Select the Wavelength. . . . . . . . . . . . . . . . . . . . . . . 3-3
Operation Overview . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3-3

SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual v
Chapter 4 Maintenance and Troubleshooting . . . . . . . . . . . . . . 4-1
Technical Support. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
Warnings and Cautions . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-3
General . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Cleaning . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-4
Cleaning 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 190 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . B-3
Index. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .I-1

vi SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
Figures Figure 1.1: SPECTRAmax 190 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1-3
Figure 1.2: Major Areas of the SPECTRAmax 190 . . . . . . . . . . . . . . . . . .1-5
Figure 1.3: Control Panel . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-5
Figure 1.4: Microplate Drawer . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-7
Figure 1.5: Components on the Back Panel of the
SPECTRAmax 190 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-8
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 Table 4.1: Error Codes. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4-8

SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual vii
Conventions
Used in this
Manual
The names of keys that appear on the SPECTRAmax 190 control panel are
shown in boxed Helvetica type. Example:
[Drawer] .
Italic and boldface type are used for emphasis. Examples: “Press carefully to
engage,”
“Do not press down.”
A note provides information that will help you properly execute an
action or procedure.
Indicates an action or condition that could potentially dam-
age the instrument or one of its components or could result in loss of
data.
Indicates a situation that could result in potential injury to a
person working with the system.
Indicates a condition involving potentially infectious bio-
logical agents requiring that proper handling precautions be taken.
Glossary of
Terms
Absorbance, A
The amount of light absorbed by a solution. To measure absorbance accu-
rately, it is necessary to eliminate light scatter. In the absence of turbidity,
absorbance = optical density.
A = log (I
0
/I)
I
0
= incident light
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
0
%T = 100 T
NOTE:
!CAUTION:
WARNING:
BIOHAZARD:

viii SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual

Chapter 1 Instrument Description
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 . . . . . . . . . . . . . . . . . . . . . .1-7
The Back Panel. . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-8
Functional Description . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-9
Stand-Alone Operation . . . . . . . . . . . . . . . . . . . . . .1-9
Wavelength Selection . . . . . . . . . . . . . . . . . . . . . . . .1-9
Temperature Regulation . . . . . . . . . . . . . . . . . . . . .1-9
Computer Control. . . . . . . . . . . . . . . . . . . . . . . . . .1-10
Specifications. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . .1-12

Chapter 1
1-2 SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual

Instrument Description
SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual 1-3
Introduction General Overview
The SPECTRAmax™ 190 microplate spectrophotometer provides rapid
and sensitive measurements of a variety of analytes across a wide range of
concentrations. It measures the optical density (OD) of samples in 96-well
microplates at a selected wavelength for a single point in time (endpoint),
over a speciÞed period of time (kinetic), or over a selected wavelength range
(spectral scan).
Figure 1.1: The SPECTRAmax 190 System
The on-board microprocessor calculates and reports the absorbance (or
% Transmittance) for each well of a microplate. Data from multiple wave-
lengths 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 DNA quantitation, endpoint assays (quantitation
of cytoproliferation by MTT reduction, colorimetric protein assays, 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-well microplates, strip wells, and Þlter-bottom microplates
can be used in the SPECTRAmax 190.
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 per-
form 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.

Chapter 1
1-4 SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
PathCheck
™
As predicted by the Beer Lambert law of light absorption, absorbance is pro-
portional 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 difÞcult
to perform extinction-based assays and confusing to compare results between
microplate readers and spectrophotometers.
This pathlength correction is accomplished only with the use of SOFTmax
PRO software. SOFTmax
®
PRO software from Molecular Devices provides
full instrument control and statistical data analysis.
The standard pathlength of a cuvette is the
conventional basis for quantifying the unique
absorptivity properties of compounds in solution.
Quantitative analyses can be performed on the
basis of extinction coefÞcients, without standard
curves (e.g., NADH-based enzyme assays). When
using a cuvette, the pathlength is known and is
independent of sample volume, so absorbance is
proportional to concentration.
In the microplate, pathlength is
dependent on the
liquid volume, so absorbance is proportional to
both the concentration and the pathlength of the
sample. Standard curves are often used to
determine analyte concentrations in vertical-beam
photometry of unknowns, yet errors can still arise
from pipetting the samples and standards. The
PathCheck feature of the SPECTRAmax 190
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 correcting the microwell
absorbance values is accurate to within 2.5% of the
values obtained directly in a 1-cm cuvette.
Horizontal
Horizontal
Light Path
Cuvette
Vertical
Vertical Light Path
Microplate Wells

Instrument Description
SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual 1-5
Component
Description
The main components of the SPECTRAmax 190 are:
• The control panel
• The microplate drawer
• The back panel (connections and power switch)
Figure 1.2: Major areas of the SPECTRAmax 190
Control Panel
Figure 1.3: Control Panel
The control panel consists of an LCD and seven pressure-sensitive, color-
coded membrane keys which can be used to perform the stand-alone func-
tionality of the SPECTRAmax 190, including setting the temperature inside
the microplate drawer, setting the wavelength at which to read the micro-
plate, 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
settings.
Control Panel
Microplate
Drawer
Back Panel

Chapter 1
1-6 SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
Keys
(Incubator) Enables/disables the incubator function.
(Incubator) Allows you to enter a set point to regulate the
microplate chamber temperature. Pressing the up or down
arrow key scrolls up or down, starting at the previous tem-
perature setting (or the default of 37.0°C, if no setting had
been made). Pressing the up (
▲ ) or down ( ▼ ) arrow once in-
crements or decrements the temperature shown in the dis-
play 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 dis-
play will not change. If you decrement the setting to the low-
est limit, 15°C, and continue to press the down (
▼ ) arrow, the
display will not change.
If the incubator is disabled, pressing the
[Temp On/Off] key
will
enable the incubator.
Selects the wavelength that will be used for reading the
microplate manually. Pressing the up or down arrow key
scrolls up or down through the available wavelengths, start-
ing at the previous 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 decrements the wavelength shown in the dis-
play by 10 nm until it is released. If you increment the set-
ting to the highest limit (850 nm) and continue pressing the
up (
▲ ) arrow, the display returns to the lowest possible set-
ting (340 nm) and begins incrementing from there. The
inverse is true for decrementing by pressing the down (
▼ )
arrow.
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.
Initiates the reading of the microplate in stand-alone mode.
Data is sent to the printer port of the SPECTRamax 190.
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 approxi-
mately 10 seconds to assist in maintaining temperature con-
trol within the microplate chamber.
!CAUTION:
NOTE:

Instrument Description
SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual 1-7
If the SPECTRAmax 190 is idle, all keys on the control panel are
active. The instrument will automatically 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 temperature of the plate drawer is shown in the
SOFTmax PRO software.) The middle of the display shows the wavelength.
The Microplate Drawer
Figure 1.4: Microplate Drawer
The microplate drawer is located on the right side of the SPECTRAmax 190
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 beep-
ing sound will alert you approximately two seconds before the drawer closes
automatically.
NOTE:
A1 Indicator

Chapter 1
1-8 SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
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 190 can accommodate standard 96-well microplates, strip
wells, and Þlter-bottom microplates.
Not all manufacturers’ microplates are the same with regard to design, mate-
rials, or conÞguration. Temperature uniformity within the microplate may
vary depending on the type of microplate used.
The Back Panel
Figure 1.5: Components on the Back Panel of the SPECTRAmax 190
The following components are located on the back panel of the SPECTRAmax
190 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 exter-
nal computer)—plug the 8-pin DIN serial cable connector into this port; the
other end attaches to the serial (modem) port of the computer.
•
Labels —provide information about the SPECTRAmax 190, such as line
voltage rating, cautionary information, serial number, etc. Record the serial
number shown on this label for use when contacting Molecular Devices
Technical Services.
NOTE:
Computer (RS-232)
Printer (Centronics)
Power Switch
Fuse Box Cover Power Cord
Receptacle
Label

Instrument Description
SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual 1-9
Functional
Description
Stand-Alone Operation
The paragraphs that follow describe functions that can be activated during
stand-alone operation.
When operating the SPECTRAmax 190 as a stand-alone system, you can
choose a temperature setting, choose a wavelength, and make an Endpoint
read. Data is sent to the printer port of the SPECTRAmax 190. The wavelength
is chosen by pressing the up or down arrows beneath the display labeled
λ .
Temperature regulation of the 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.
Wavelength Selection
The display on the control panel shows the currently selected measurement
wavelength (
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 190 has been designed to regulate the temperature of the
microplate chamber from 4°C above ambient to 45°C. Upon powerup, when
the incubator is off, the temperature in the SPECTRAmax 190 microplate
chamber is ambient and isothermal. Pressing the incubator
[TEMP On/Off]
key will cause the instrument to begin warming the microplate chamber.
The temperature set point defaults to 37.0°C at start-up.
Accuracy of the temperature set point is guaranteed only 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 regula-
tion off. The microplate drawer will open and the temperature within the
chamber will begin returning to ambient.
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 within the microplate chamber is achieved through
electric heaters, a fan, efÞcient insulation, and temperature sensors. The heat-
ers are located in the chamber which is insulated to maintain the temperature
NOTE:
NOTE:

Chapter 1
1-10 SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
set point. The sensors, also mounted inside the chamber, measure the air tem-
perature. 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 temper-
ature variation of the air inside the chamber of less than 1.0°C. (The tempera-
ture uniformity within the microplate will depend upon the design, materials,
and/or conÞguration of that component.)
Computer Control
The SPECTRAmax 190 is equipped with an 8-pin DIN RS-232 serial port
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 190.
SOFTmax PRO makes it possible to access the full capabilities of the
SPECTRAmax 190. 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 850-nm
range
- Extend kinetic run times up to 99 hours
- 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
• Use PathCheck to normalize the absorbance readings in each microplate
well to a 1-cm pathlength
• Design microplate templates to simplify data reduction
- Identify groups of wells with labels of your choice
- Identify individual wells with unique names
- Blank the entire plate, groups, and/or individual wells
• Save instrument settings, template formats, and data analysis parameters
for microplates as assay protocol Þles and recall them for later use
- Rapid instrument and analysis setup for repeated microplate assays
- Uniform analysis for equivalent microplates
•Turn the incubator on or off to control the temperature in the microplate
drawer

Instrument Description
SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual 1-11
•Acquire data from the SPECTRAmax 190
- Save data files for in-depth analysis at a later time
- Save multiple microplates with individual template and data analysis pa-
rameters 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
-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 lim-
its in a microplate format
- Kinetic or spectrum plots of all 96 microplate wells
- Enlarge the display of individual well plots and overlay multiple well
plots
• 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 ex-
periments
• 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 data-
base programs
For a complete description of the features of SOFTmax PRO, refer to the
SOFTmax PRO User’s Manual.

Chapter 1
1-12 SPECTRAmax 190 Microplate Spectrophotometer Operator’s Manual
SpeciÞcations Thermal speciÞcations for microplates used in the SPECTRAmax 190 apply to
ßat-bottom microplates with isolated wells. All other microplate speciÞca-
tions apply to standard 96-well polystyrene ßat-bottom microplates.
Technical speciÞcations are subject to change without notice.
Photometric Performance
Wavelength range 190–850 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
Photometric accuracy/linearity
(microplate), 0–2.0 OD: 190–850 nm <± 1.0% and ± 0.006 OD
Photometric precision
(repeatability), 0–2.0 OD: 190–850 nm <± 1.0% and ± 0.003 OD
Stray light ≤0.05% at 340 nm
Photometric stabilization Instantaneous
Photometric drift None—continuous referencing of monochromatic output
Calibration Automatic before first kinetic read and before every end-
point reading
Optical alignment None required
Light source Xenon flash lamp (5 watts)
Average lamp lifetime 1 billion flashes
Illumination Top down
Photodetectors Silicon photodiode
Photometric Analysis Modes
Stand-Alone
•Single wavelength Absorbance reading of the microplate
Using SOFTmax PRO
• Express data as Absorbance or %Transmittance
• Single wavelength reading of microplate
•
Multiple wavelength (up to six) reading of microplate
• Kinetic and kinetic graphics of microplate
• Spectral scan (190–850 nm) of microplate
Measurement Time (calibration off)
NOTE: