Molecular Devices SpectraMax L User manual

0112-0174 C
October 2018
SpectraMax® L
Luminescence Microplate Reader
User Guide

SpectraMax L Luminescence Microplate Reader User Guide
2 0112-0174 C
This document is provided to customers who have purchased Molecular Devices equipment,
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Contents
Safety Information 4
Warnings, Cautions, Notes, and Tips 4
Chapter 1: Introduction 7
Instrument Features 8
Computer Integration 9
PMT Technology 9
Chapter 2: Setting Up the Instrument 11
Unpacking the Instrument 15
Assembling the Injectors 17
Installing the Filters 19
Installing the Barcode Reader 21
Installing the Software 21
Changing the Plate Aperture 21
Chapter 3: Getting Started 23
Software Ribbon 23
Instrument Calibration 24
Chapter 4: Protocols 26
Temperature Regulation 26
Injector Protocols 26
Protocol Settings 28
Powering Off the Instrument 31
Chapter 5: Maintenance 32
Cleaning the Instrument 32
Replacing Injector Tips 33
Replacing Injector Tubing 36
Chapter 6: Before You Move the Instrument 41
Packing the Instrument 42
Chapter 7: Opening the Drawer Manually 43
Obtaining Support 44
Appendix A: Instrument Specifications 45
Appendix B: Electromagnetic Compatibility 46

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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.
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SpectraMax L Luminescence Microplate Reader User Guide
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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.
WARNING! HIGH VOLTAGE. Within the instrument is the potential of an
electrical shock hazard existing from a high voltage source. Read and
understand all safety instructions before you install, maintain, and service the
instrument.
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 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.
Never try to physically restrict the moving components of the instrument.
WARNING! Do not attempt to access the interior of the instrument unless
specifically instructed to do so. The moving parts inside the instrument can
cause injury. Do not operate the instrument with any covers or panels
removed.
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.

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Chapter 1: Introduction
The extreme flexibility and high sensitivity of the SpectraMax L instrument makes it ideal
for applications within the fields of biochemistry, cell biology, immunology, molecular
biology, and microbiology. Typical applications include luciferase reporter gene, ATP,
Ca2+, aequorin, and ELISAs.
The instrument is a high-performance microplate reader that measures luminescence in
96-well and 384-well plates.
There are multiple instrument configurations that enable you to vary the components
that are included in the instrument when it is shipped to you from the factory.
Base - one PMT, manual aperture
Injectors - up to four injectors
PMTs - one or two PMTs
Dual Wavelength
Automatic Aperture
Filters
Barcode Reader
In the SoftMax® Pro Software, select the Home tab and click Instrument Configuration to
see the configuration of your instrument.
Read Modes
The components included in your instrument configuration determine which read modes
are available:
Lumi (Luminescence): For all instrument configurations, you can select the Lumi read
mode. When the instrument has two PMTs and no filters, the instrument runs Lumi
protocols that use both PMTs to read 2 wells simultaneously and it assumes the
readings of the PMTs are comparable.
Lumi2W (Luminescence Dual Wavelength): When the instrument has two PMTs and
the PMTs are equipped with filters, you can select the Lumi2W read mode to use the
PMTs one at a time to detect the two different wavelengths.
Read Types
For all instrument configurations, you can select the following read types:
Endpoint: A read of each well is taken in the center of each well, either at all
wavelengths if filters are not used, or at specified wavelengths if filters are used. Raw
data values are reported as relative light units (RLU). Each well can receive up to two
injections and then is read once. You can set a delay time after each injection, from
0.1 to 3,600 seconds for the P-injection, and from 0 to 3,600 seconds for the M-
injection.
Dual Read: Each well receives two injections, each followed by a read. The first
injection is the P-injection with a subsequent read. The second injection is the M-
injection with a subsequent read. You can set a delay time between injection and
read, from 0.1 to 3,600 seconds for the P-injection, and from 0 to 3,600 seconds for
the M-injection.
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Kinetic: The instrument collects data over time with multiple reads taken in the
center of each well at regular intervals. Each well can receive multiple injections and
reads. You can specify single injections, followed by single reads, for multiple time
points. The minimum integration interval is dependent on the number of wells you
select, the integration time you select, and the number of injections you select.
Fast Kinetic: The instrument performs repeated reads of each well up to a 200 point
maximum integration. All reads of a single well are made before the next well is read.
Pre-reading a plate is not possible in Fast Kinetic read type. Each well can receive
multiple injections and reads. You can specify multiple injections and multiple reads,
for multiple times. Integration time can be set from 0.01 to 3600 seconds.
When the instrument configuration includes the variable-volume injectors option, it can
measure both flash and glow luminescence. High-precision peristaltic pumps ensure
precise injection volumes and rapid sample mixing. Continuous monitoring between
injection and measurement provides for extremely fast reactions and reduced delay time.
96-well Standard plates - 10 µL to 200 µL
384-well and 96-well half area plates - 10 µL to 80 µL
Optional multiple photomultiplier tubes (PMTs) and multiple injector configurations
enable increased throughput and wavelength-dependent assays such as BRET1 and
BRET2.
Instrument Features
The instrument can have the following features:
Greater than nine orders of dynamic range
Reads both 96-well and 384-well plates
Dual injection into both 96-well and 384-well plates
Programmable injection speeds
AutoWash flush of tubing and tips when you power off the instrument
StakMax® Microplate Handling System and robot compatible
Landscape and portrait plate orientation
Optional dual PMT configuration with filter positions for wavelength dependent
assays such as BRET
Optional dual PMT configuration for higher throughput
External injector module for dual-PMT, four injector configurations
Isothermal temperature control in the plate chamber
SoftMax Pro Software
Optional automatic aperture switching between 96-well and 384-well plates
Optional barcode reader to read labels on plates
The instrument can accommodate SBS-standard 96-well and 384-well plates with
dimensions of 128.2 mm long, 86.0 mm wide, and heights from 14 mm to 15 mm.

Chapter 1: Introduction
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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.
PMT Technology
The instrument uses high-sensitivity, low-noise PMTs to detect emitted luminescence.
PMTs convert incident photons into electrons through the photoelectric effect. When an
incident photon impinges on the active surface of the PMT (the photocathode), an
electron is generated. The electron flows through a series of electron multipliers
(dynodes) to the anode. The amount of current that flows from the anode is directly
proportional to the number of photons at the photocathode.
The amount of amplification that a PMT can produce depends on the number of dynodes
in the PMT, and the voltage that is applied to it.
The PMTs in the instrument meet two essential criteria for high-quality detection. First,
since different photocathodes have varying sensitivities to specific wavelengths of light
(quantum efficiency), the PMTs have been optimized for the typical wavelengths of
luminescent emission light. Second, the PMTs have exceptional reliability and optimal
signal-to-noise performance.

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PMT Modes
The instrument can read PMT values in the following modes:
Mode Recommended Use Benefits Dynamic
Range
(RLU)
Target
Calibration
Wavelength
Required
Analog
Only
Very bright assays. Uses low PMT voltage
to prevent PMT
saturation
1 x 107
to
1 x 109
Yes
Photon
Counting
Assays requiring high sensitivity with
an average dynamic range. Use when
non-calibrated data is desired.
Excellent discrimination
between signal and
background noise
10 to
25 x 106
No
AutoRange Assays requiring high sensitivity with
a wide dynamic range.
High sensitivity and
wide dynamic range
10 to
2 x 108
Yes
MaxRange Assays with massive dynamic range
that can tolerate sub-second delays if
the signal is saturated.
Widest dynamic range
and high sensitivity
10 to
1 x 109
Yes
Operating Modes

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Chapter 2: Setting Up the Instrument
Before you unpack and set up the SpectraMax L Microplate Reader instrument, prepare a
dry, flat work area away from direct sunlight, dust, drafts, vibration, and moisture.
Remember to leave sufficient space for the instrument, computer, and required cables.
See Instrument Specifications on page 45.
The package contains the instrument plus one or more boxes that contain accessories.
Illustration Part Number Description
SoftMax® Pro Software Latest
Version
SoftMax Pro Software installation guide with Product
Key
5059573 Instrument installation guide
4400-0002 or
4400-0036
Region Specific Power cord, 115 V
or Power cord, 230 V
4200-0076 Power supply adapter
0700-0861
1-2100-0926
USB cable, 2meter (6.56foot)
8500-1229 Drawer pull tool
5060689 Flat head screw driver
Package Contents for Base Instrument
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Illustration Part Number Description
8500-1559 M2 Hex Wrench
9000-0133 SpectraMax® Mouse Pad
9000-0972 Package of 5 sample microplates
2400-2464 Dust Cover
5000505 CE Declaration of Conformity
5019680 Card, User Guide Download
0120-1319 Datasheet, Innovative Solutions
0120-1437 Datasheet, StackMax Microplate Handling System
0120-1477 Datasheet, MicroplateReader Validation and Compliance
Package Contents for Base Instrument Continued
For a complete list of the contents of the package, see the enclosed packing list.

Chapter 2: Setting Up the Instrument
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Illustration Part Number Description
4400-0002 or
4400-0036
Injector module
Power cord, 115 V
or Power cord, 230 V
4200-0900 Injector module
Power supply adapter
0310-5238 Waste drain hose
0700-0861 Injector module communication cable
9000-0964 2 Reagent bottles, amber
9000-0965 2 Wash bottles, natural
8500-1072 Wrench, Fitting, Upchurch
5004193 4 Set screws, 1/4-28 x 1/4", oval point
Additional Package Contents for Other Instrument Configurations

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14 0112-0174 C
Illustration Part Number Description
0310-5705 Tip extractor
5000592 Diagram, Assembly, Canopy and Tray
0310-5677 Barcode reader
Additional Package Contents for Other Instrument Configurations (continued)
For a complete list of the contents of the package, see the enclosed packing list.

Chapter 2: Setting Up the Instrument
0112-0174 C 15
Unpacking the Instrument
The packaging is designed to protect the instrument during transportation. Retain the
container and the packing materials for future transport needs. Do not use tools that can
damage the packaging or the instrument.
CAUTION! When transporting the instrument, warranty claims are void if
improper packing results in damage to the instrument.
The main instrument components are:
Item Name Description
1 Top cover Cover for base configuration
2 Plate drawer Houses the plate in landscape or portrait orientation
3 Power switch Powers the instrument on and off
4 Control panel Contains the instrument status lights and the Drawer button
5 Injector ports Two-injector configuration
6 Injector module Factory-installed injector module four-injector configuration
Instrument Components
WARNING! LIFTING HAZARD. To prevent injury, use a minimum of two
people to lift the instrument.

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CAUTION! Do not touch or loosen any screws or parts other than those
specifically described in the instructions. Doing so may cause misalignment and
voids the instrument warranty.
To unpack the instrument:
1. Check the box for visible damage that occurred during transportation. In case of
damage, inform the supplier immediately and keep the damaged packaging.
CAUTION! Keep the box upright. Do not tip or tilt the box or place it on its
side.
2. With the box facing up as indicated on the package, use a box cutter to carefully cut
open the top of the box.
3. Remove the accessory boxes and lift the cardboard and foam packing materials
above the instrument.
4. Lift the instrument, along with the plastic bag, up and out of the shipping box.
5. Remove the plastic bag and place the instrument on a level surface, near the
controlling computer, with the back panel facing you.
Item Name
1 Electrical power port
2 Computer communication USB port
3 Injector module communication port
4 Injector waste port
Back Panel
6. Insert the USB cable in the USB port on the back of the instrument and insert the
other end in the USB port on the control computer.
7. Insert one end of the instrument power supply into the instrument power cord port
and the other end into the power cord. Then plug the power cord into a grounded
electrical outlet.

Chapter 2: Setting Up the Instrument
0112-0174 C 17
8. If the instrument configuration includes an injector module, insert one end of the
injector module power supply into the injector module power port on the back of the
injector module and the other end into the power cord. Then plug the power cord
into a grounded electrical outlet.
9. Insert the square end (release tab up) of the injector module communication cable
into the PORT on the back panel of the instrument and then insert the round end
into the COMMIN port on the rear of the injector module.
Item Name
1 PORT - Injector module communication port on instrument
2 COMM IN - Communication in port on injector module
Communication Cable Connection for Injector Module
10. Turn the instrument around so that the control panel faces you. Ensure no cables run
beneath the instrument. Leave sufficient space between the back of the instrument
and the nearest objects or surfaces to enable the ability to lift the instrument hood
and injector hood and to ensure proper ventilation and cooling.
Assembling the Injectors
The base instrument configuration does not include injectors. The instrument
configuration can have several factory installed injector setups. For all instrument
configurations that include injectors, the wash bottles and reagent bottles remain
outside the instrument.
When the instrument configuration includes two injectors there is one pre-injector (P-
injector) and one measurement injector (M-injector). The injectors are built into the
instrument with injector ports located on the left side of the instrument.

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Item Name Description
1 P-
Reagent
Connection for tubing from the P-reagent bottle. Controlled by P-injector commands in the
SoftMax Pro Software. If plate settings include both a P-injection and a M-injection, the P-
injection occurs before the M-injection.
2 M-
Reagent
Connection for tubing from the M-reagent bottle. Controlled by M-injector commands.
3 P-Wash Connection for tubing from P-wash bottle. Washes the P-injector from injector port
through injector tip. Controlled by the P-Wash commands and automatically when you turn
the instrument off.
4 M-
Wash
Connection for tubing from M-wash bottle. Washes the M-injector from injector port
through injector tip. Controlled by the M-Wash commands and automatically when you
turn the instrument off.
Injector Ports - Base Configuration Instrument
When the instrument configuration includes four injectors, the instrument has a factory-
installed injector module on the left side. The injector module tubing is grouped into P-
reagent, M-reagent, P-wash, and M-wash bundles. Injector module tubing exits the
injector module through the rear of the instrument.
1. Before you connect the tubing, clean the wash bottles with an appropriate solvent.
2. For the base instrument configuration, connect the tubing to the P-Wash injector
port on the instrument.
3. For the base instrument configuration, connect the tubing to the M-Wash injector
port on the instrument.
4. For the base instrument configuration, connect one piece of tubing to the P-Reagent
port and the other to the M-Reagent port on the instrument.
5. Connect the waste line drain hose to the Waste port on the instrument back panel.
Place the other end of the waste line in a suitable waste container.
Note: Make sure the waste container is in a secure location and can not spill.

Chapter 2: Setting Up the Instrument
0112-0174 C 19
Installing the Filters
If you intend to run wavelength-dependent assays, perform the following steps to install
filters in the instrument.
Required tools:
M2 hex wrench
Slotted screwdriver
Powder-free latex or nitrile gloves
To install the filters:
1. Power off the instrument.
2. Loosen the captive screws on the front of the instrument and then raise the top
cover up and over the instrument.
3. Remove the two hex screws that secure each detector to the instrument chassis. The
screws are located inside the access holes in the detector adapter.
CAUTION! You must keep the PMT away from any direct light source. Light
can damage the PMT and cause temporarily high background reads.
Item Name
1 Access holes for screws
2 Detector
3 Detector adapter
Detector Attached to Instrument Chassis
4. Pull the detector away from the instrument chassis and then place it on a clean
surface away from any direct source of light.

SpectraMax L Luminescence Microplate Reader User Guide
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5. Loosen the two flat head screws that secure the filter cover plate to the detector.
Item Name
1 Detector
2 Filter cover plate
3 Flat head screws
4 Detector adapter
Detector With Filter Cover Plate Attached
6. Lift the filter cover plate off the detector and then remove the filter (or silver open
ring) from inside the detector.
7. Insert the new filter into the detector, position the filter cover plate on the detector,
and then tighten the captive screws.
8. Position the detector on the instrument chassis and then replace the screws that
secure the detector to the instrument chassis.
9. Lower the top cover and then tighten the captive screws on the front of the
instrument.