Molecular Devices EMax Plus Manual de usuario

5032018 C

October 2018

EMax®Plus

Microplate Reader

User Guide

EMax Plus Microplate Reader User Guide

2 5032018 C

This document is provided to customers who have purchased Molecular Devices equipment, software,

reagents, and consumables to use in the operation of such Molecular Devices equipment, software,

reagents, and consumables. This document is copyright protected and any reproduction of this

document, in whole or any part, is strictly prohibited, except as Molecular Devices may authorize in

writing.

Software that may be described in this document is furnished under a non-transferrable license. It is

against the law to copy, modify, or distribute the software on any medium, except as specifically

allowed in the license agreement. Furthermore, the license agreement may prohibit the software

from being disassembled, reverse engineered, or decompiled for any purpose.

Portions of this document may make reference to other manufacturers and/or their products, which

may contain parts whose names are registered as trademarks and/or function as trademarks of their

respective owners. Any such usage is intended only to designate those manufacturers’ products as

supplied by Molecular Devices for incorporation into its equipment and does not imply any right and/or

license to use or permit others to use such manufacturers’ and/or their product names as trademarks.

Each product is shipped with documentation stating specifications and other technical information.

Molecular Devices products are warranted to meet the stated specifications. Molecular Devices makes

no other warranties or representations express or implied, including but not limited to, the fitness of

this product for any particular purpose and assumes no responsibility or contingent liability, including

indirect or consequential damages, for any use to which the purchaser may put the equipment

described herein, or for any adverse circumstances arising therefrom. The sole obligation of Molecular

Devices and the customer's sole remedy are limited to repair or replacement of the product in the

event that the product fails to do as warranted.

For research use only. Not for use in diagnostic procedures.

The trademarks mentioned herein are the property of Molecular Devices, LLC or their respective owners. These trademarks may not

be used in any type of promotion or advertising without the prior written permission of Molecular Devices, LLC.

Patents: http://www.moleculardevices.com/patents

Product manufactured by Molecular Devices, LLC.

3860 N. First Street, San Jose, California, 95134, United States of America.

Molecular Devices, LLC is ISO 9001 registered.

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Contents

Safety Information 4

Warnings, Cautions, Notes, and Tips 4

Chapter 1: Introduction 8

Computer Integration 8

Absorbance Read Mode 9

Endpoint Read Type 10

Kinetic Read Type 10

Chapter 2: Setting Up the Instrument 11

Connecting Instrument Cables 12

Chapter 3: Getting Started 13

Selecting the Instrument in the Software 13

SoftMax Pro Protocol Settings 14

Loading and Unloading Plates 15

Chapter 4: Maintenance 17

Cleaning the Instrument 17

Replacing the Halogen Lamp 18

Adjusting the Lamp 20

Obtaining Support 21

Appendix A: Instrument Specifications 22

Physical Specifications 23

Electromagnetic Compatibility 23

System Dimensions 24

EMax Plus Microplate Reader User Guide

4 5032018 C

Safety Information

Information about the safe use of the instrument from Molecular Devices® includes an

understanding of the user-attention statements in this guide, the safety labels on the

instrument, precautions to follow before you operate the instrument, and precautions to

follow while you operate the instrument.

Make sure that everyone involved with the operation of the instrument has:

Received instruction in general safety practices for laboratories.

Received instruction in specific safety practices for the instrument.

Read and understood all Safety Data Sheets (SDS) for all materials being used.

Read and observe all warnings, cautions, and instructions. The most important key to safety

is to operate the instrument with care.

WARNING! If the instrument is used in a manner not specified by Molecular

Devices, the protection provided by the equipment might be impaired.

Warnings, Cautions, Notes, and Tips

All warning symbols are framed within a yellow triangle. An exclamation mark is used for

most warnings. Other symbols can warn of other types of hazards such as biohazard,

electrical, or laser safety warnings as are described in the text of the warning. Follow the

related safety information.

The following user attention statements might be displayed in the text of Molecular Devices

user documentation. Each statement implies the amount of observation or recommended

procedure.

WARNING! A warning indicates a situation or operation that could cause

personal injury if precautions are not followed.

CAUTION! A caution indicates a situation or operation that could cause damage to

the instrument or loss of data if correct procedures are not followed.

Note: A note calls attention to significant information.

Tip: A tip provides useful information or a shortcut, but is not essential to the

completion of a procedure.

Safety Information

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Symbols on the Instrument

Each safety label found on the instrument contains an alert symbol that indicates the type of

potential safety hazard.

Symbol Indication

Consult the product documentation.

Potential biohazard.

Power switch indicates power on.

Power switch indicates power off.

Required in accordance with the Waste Electrical and Electronic Equipment (WEEE)

Directive of the European Union. It indicates that you must not discard this electrical or

electronic product or its components in domestic household waste or in the municipal

waste collection system.

For products under the requirement of the WEEE directive, contact your dealer or

local Molecular Devices office for the procedures to facilitate the proper collection,

treatment, recovery, recycling, and safe disposal of the device.

- - California proposition 65 requires businesses to provide

warnings to Californians about significant exposures to chemicals that cause cancer, birth

defects, or other reproductive harm.

EMax Plus Microplate Reader User Guide

6 5032018 C

Electrical Safety

To prevent electrical injuries and property damage, inspect all electrical equipment before

use and report all electrical deficiencies. Contact Molecular Devices technical support for

equipment service that requires the removal of covers or panels.

To prevent electrical shock, use the supplied power cord and connect to a properly

grounded wall outlet.

To ensure sufficient ventilation and provide access to disconnect power from the

instrument, maintain a 20cm to 30cm (7.9in. to 11.8in.) gap between the rear of the

instrument and the wall.

Power off the instrument when not in use.

Chemical and Biological Safety

Normal operation of the instrument can involve the use of materials that are toxic,

flammable, or otherwise biologically harmful. When you use such materials, observe the

following precautions:

Handle infectious samples based on good laboratory procedures and methods to

prevent the spread of disease.

Observe all cautionary information printed on the original containers of solutions before

their use.

Dispose of all waste solutions based on the waste disposal procedures of your facility.

Operate the instrument in accordance with the instructions outlined in this guide, and

take all the required precautions when using pathological, toxic, or radioactive materials.

Splashing of liquids can occur. Take applicable safety precautions, such as using safety

glasses and wearing protective clothing, when working with potentially hazardous

liquids.

Observe the applicable cautionary procedures as defined by your safety officer when

using hazardous materials, flammable solvents, toxic, pathological, or radioactive

materials in or near a powered-up instrument.

WARNING! Never use the instrument in an environment where potentially

damaging liquids or gases are present.

Safety Information

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Moving Parts Safety

The plate carrier on the EMax Plus moves in and out of the read chamber during the read,

and then returns to the home position after the read ends. If the plate carrier does not

return to the home position, select the Operations tab and click Initialize.

The instrument contains moving parts that can cause injury. Under normal conditions, the

instrument is designed to protect you from these moving parts.

WARNING! If the instrument is used in a manner not specified by Molecular

Devices, the protection provided by the equipment might be impaired.

To prevent injury:

Never try to exchange labware, reagents, or tools while the instrument is operating.

The instrument calibrates the lamp before moving the microplate carrier into the

chamber for each read. Between multiple-wavelength or kinetic reads, the carrier waits in

the home position before the read continues. Do not try to remove the plate until the

software indicates that the entire read is complete.

Never try to physically restrict the moving components of the instrument.

Keep the instrument work area clear to prevent obstruction of the movement.

CAUTION! To prevent damage to the instrument, the height of the plate must not

exceed 15mm.

Note: Observe all warnings and cautions listed for all external devices attached to or in

use during the operation of the instrument. See the applicable user guide for the

operating and safety procedures of that device.

CAUTION! Never touch the optic mirrors, lenses, filters, or cables. The optics are

extremely delicate, and critical to the function of the instrument.

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Chapter 1: Introduction

The EMax®Plus Microplate Reader is a filter-based absorbance microplate reader with eight

standard filters for a wide range of applications.

The instrument can read 96-well plates to run protocols with the visible Absorbance (ABS)

read mode with the Endpoint and Kinetic read types.

The high sensitivity and flexibility of the EMax Plus Microplate Reader make it useful for

applications in the fields of biochemistry, cell biology, immunology, molecular biology, and

microbiology.

Typical applications include ELISA, total protein, and cell proliferation.

Application notes with specific application protocol suggestions are in the Information

Center and Knowledge Base on the Molecular Devices web site at

www.moleculardevices.com.

Computer Integration

Each Molecular Devices microplate reader is shipped with a license key for the SoftMax® Pro

Data Acquisition and Analysis Software that you install on the computer that you use to

operate the instrument. The SoftMax Pro Software provides integrated instrument control,

data display, and statistical data analysis.

You should install the SoftMax Pro Software on the computer before you set up the

instrument. Please be aware that some updates to the SoftMax Pro Software require a

purchase. Contact Molecular Devices before you update the software. To download the

latest version of the software, visit:

https://www.moleculardevices.com/products/microplate-readers/acquisition-and-analysis-

software/softmax-pro-software#Order.

Note: For information about the computer specifications that are required to run the

software, the software installation and licensing instructions, and the directions to

create the software connection between the computer and the instrument, see the

SoftMax Pro Data Acquisition and Analysis Software Installation Guide.

To prevent data loss, turn off all sleep and hibernation settings for the hard disk, the CPU,

and the USB ports. Disable automatic Windows updates. Update Windows manually when

you do not use the computer to control an instrument. You can set these options in

Windows Control Panel.

You can connect the instrument to a printer and operate the instrument in stand alone

mode to run basic Absorbance read mode Endpoint read type protocols. In stand alone

mode, the instrument control panel enables you to adjust the temperature and the

wavelength to do fixed point plate reads. For the SpectraMax Plus 384 you can do fixed point

cuvette reads. Stand alone mode is not available for the VersaMax.

EMax Plus Microplate Reader User Guide

9 5032018 C

To run protocols that require advanced acquisition settings or to run Absorbance read mode

Kinetic read type and Spectrum read type protocols, you must connect the instrument to a

computer and use the SoftMax Pro Software to operate the instrument.

Absorbance Read Mode

The instrument uses the Absorbance (ABS) read mode to measure the Optical Density (OD)

of the sample solutions.

Absorbance is the quantity of light absorbed by a solution. To measure absorbance

accurately, it is necessary to eliminate light scatter. If there is no turbidity, then

absorbance=optical density.

A=log10(I0/I)=–log10(I/I0)

where I0is intensity of the incident light before it enters the sample divided by the light after

it passes through the sample, and Ais the measured absorbance.

When you measure absorbance, the pathlength is defined by the volume of sample in the

well. To make sure that all the wells in the plate contain the same volume, use an accurate

liquid handler to dispense the sample into the plate.

The temperature-independent PathCheck® Pathlength Measurement Technology

normalizes your absorbance values to a 1cm path length based on the near-infrared

absorbance of water.

The instrument enables you to choose whether to display absorbance data as Optical

Density (OD) or %Transmittance (%T) in the Reduction dialog.

Optical Density

Optical density (OD)is the quantity of light passing through a sample to a detector relative to

the total quantity of light available. Optical Density includes absorbance of the sample plus

light scatter from turbidity and background. You can compensate for background using

blanks.

A blank well contains everything used with the sample wells except the chromophore and

sample-specific compounds. Do not use an empty well for a blank.

Some applications are designed for turbid samples, such as algae or other micro-organisms

in suspension. The reported OD values for turbid samples are likely to be different when read

by different instruments.

For optimal results, you should run replicates for all blanks, controls, and samples. In this

case, the blank value that will be subtracted is the average value of all blanks.

% Transmittance

%Transmittance is the ratio of transmitted light to the incident light for absorbance reads.

T=I/I0

%T=100T

where Iis the intensity of light after it passes through the sample and I0is incident light

before it enters the sample.

Optical Density and %Transmittance are related by the following formulas:

%T=102–OD

Chapter 1: Introduction

5032018 C 10

OD=2–log10(%T)

The factor of two comes from the fact that %T is expressed as a percent of the transmitted

light and log10(100)=2.

When in %Transmittance analysis mode, the instrument converts the raw OD values

reported by the instrument to %Transmittance using the above formula. All subsequent

calculations are done on the converted numbers.

Applications of Absorbance

Absorbance-based detection is commonly used to evaluate changes in color or turbidity,

permitting widespread use including ELISAs, protein quantitation, endotoxin assays, and

cytotoxicity assays.

Endpoint Read Type

For the Endpoint read type, a read of each plate well is taken in the center of each well, at a

single wavelength or at multiple wavelengths.

Kinetic Read Type

For the Kinetic read type, the instrument collects data over time with reads taken in the

center of each well at regular intervals.

The values calculated based on raw kinetic data include VMax, VMax per Sec, Time to VMax,

Onset Time, and more. Kinetic reads can be single-wavelength or multiple-wavelength reads.

Kinetic analysis can collect data points in time intervals of seconds, minutes, hours, or days.

Kinetic analysis has many advantages when determining the relative activity of an enzyme in

different types of plate assays, including ELISAs and the purification and characterization of

enzymes and enzyme conjugates. Kinetic analysis can provide improved dynamic range,

precision, and sensitivity relative to endpoint analysis.

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