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SpectraMax iD3s and iD5e – Multi-Mode Microplate Readers

Tunable wavelength selection, onboard injectors, and optional environmental controls to support cell-based & microbial assays

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Designed for today’s assays and tomorrow’s breakthroughs—flexible detection that evolves with your science.

The SpectraMax® iD3s and iD5e Multi-Mode Microplate Readers deliver flexible, scalable solutions for absorbance, fluorescence, and luminescence assays—with advanced capabilities for evolving research needs. The iD3s reader offers a streamlined, affordable platform with a tunable monochromator and temperature control, while the iD5e reader adds enhanced detection modes including time-resolved fluorescence (TRF) and tunable fluorescence polarization (FP) and can be expanded to include TR-FRET, HTRF, BRET and western blot. Both readers support on board fluidics with optional dual injectors.

For live-cell assays and microbial growth experiments, environmental control for CO₂/O₂ regulation and advanced shaking are available to maintain ideal growth conditions. Data acquisition and analysis are driven by SoftMax® Pro Software—the most cited microplate reader software in the industry.

Supports diverse assay needs

Perform ELISA, nucleic acid and protein quantitation, cell viability, microbial assays, western blot, and more.

Simplified setup & analysis

Over 200 preconfigured protocols and intuitive setup tools help streamline data acquisition and analysis—backed by the industry’s most cited microplate reader software.

Fast, consistent mixing for high-performance flash assays

Dual injectors available for fast kinetic assays with SmartInject® Technology, ensuring complete reagent mixing in every well for high-precision with 10 µL dead volume.

SpectraMax iD3s Multi-Mode Microplate Reader SpectraMax iD5e Multi-Mode Microplate Reader
Read modes Absorbance

Fluorescence (top/bottom read)

Luminescence (top read)

 Absorbance

Fluorescence (top/bottom read)

Luminescence (top/bottom read)

Time-Resolved Fluorescence (top/bottom read)

Fluorescence Polarization (top/bottom read) HTRF (top/bottom read) HTRF
Wavelength ranges Abs: 230 – 1000 nm

FL Ex: 250 – 830 nm

FL Em: 270 – 850 nm

Lumi: 300 – 850 nm

 Abs: 230 – 1000 nm

FL Ex: 250 – 830 nm

FL Em: 270 – 850 nm

Lumi: 300 – 850 nm

TRF Ex: 350 nm

TRF Em: 450 – 750 nm

FP: 300 – 750 nm
Wavelength selection Monochromator tunable in 1-nm increments Hybrid filters and monochromator tunable in 1 nm increments
Absorbance photometric accuracy/linearity < ±0.010 OD ±1.0%, 0 – 3 OD VIS 0 – 3 OD UV < ±0.010 OD ±1.0%, 0 – 3 OD VIS 0 – 3 OD UV
Absorbance photometric precision/repeatability < ±0.003 OD ±1.0%, 0 – 3 OD VIS 0 – 3 OD UV < ±0.003 OD ±1.0%, 0 – 3 OD VIS 0 – 3 OD UV
Sensitivity top read optimized FL: 1 pM fluoresein 96 well, 1 pM fluorescein 384 well Lumi: 2 pM ATP 96 well, 4 pM FL: Mono: 1 pM fluorescein 96 well, 1 pM fluorescein 384 well; Filters: 0.3 pM fluoresein 96 well, 0.5 pM fluorescein 384 well

Lumi: 2 pM ATP 96 well, 4 pM ATP 384 well TRF (filters with enhanced TRF module): 10 fM Europium (2 amol/well) 96 well, 10 fM Europium (1 amol/well) 384 well

FP: Mono: 2 mP @ 10nM 96 well, 2 mP @ 10nM 384 well; Filters: 1 mP @ 1 nM 96 well, 2 mP @ 1 nM 384 well
Light source(s) High power xenon flash lamp High power xenon flash lamp
Detector(s) Silicon photodiode

Ultra-cooled photomultiplier tube

 Silicon photodiode

Ultra-cooled photomultiplier tube
Read types (endpoint, kinetic, etc) Endpoint

Kinetic

Spectrum scan

Well scan

 Endpoint

Kinetic

Spectrum scan

Well scan
Read times Abs: 28 seconds

FL: 21 seconds

Lumi: 21 seconds

 Abs: 28 seconds

FL: 21 seconds

Lumi: 21 seconds

TRF: 25 seconds

FP: 35 seconds
Plate shaking Linear

Orbital

Double orbital

 Linear

Orbital

Double orbital
Injectors true true
Western blot false true
Stand-alone touchscreen operation true true
Robotics / automation true true
Plate type(s) 6-384 well plates 6-384 well plates
Temperature control Ambient + 5°C to 66°C Ambient + 5°C to 66°C
Dimensions H 40.1 cm x W 53.2 cm x D 59.8 cm H 40.1 cm x W 53.2 cm x D 59.8 cm
Validation tools Absorbance validation plate

Fluorescence validation plate

Luminescence validation plate

SoftMax® Pro GxP Software

 Absorbance validation plate

Fluorescence validation plate

Luminescence validation plate

Multi-mode validation plate

SoftMax® Pro GxP Software
SpectraMax aer gas mixer (optional) O2 control range: 1-21% in 0.1% increments

CO2 control range: 0.1-15% in 0.1% increments

O2 accuracy: ±1.0% (37°C, 1% O2)

CO2 accuracy: ±1.0% (37°C, 5% CO2)

 O2 control range: 1-21% in 0.1% increments

CO2 control range: 0.1-15% in 0.1% increments

O2 accuracy: ±1.0% (37°C, 1% O2)

CO2 accuracy: ±1.0% (37°C, 5% CO2)

Superior hybrid optical system

The high-efficiency monochromator and filter-based optical system, combined with an ultra-cooled photomultiplier tube (PMT), minimizes noise while increasing sensitivity, selectivity, and dynamic range.

Live cell assays

Advanced shaking and user-installable gas mixer (CO2/O2) that support ideal environments for cell-based and microbial growth experiments are available as optional upgrades.

Automatic filter identification

Automatically identify filter sets using NFC functionality, simplifying workflows and ensuring accuracy and reliability.

Flexible temperature control

Precise and intuitive temperature regulation—from 5°C above ambient conditions up to 66 °C—empowering your lab to perform a wide range of temperature-sensitive assays with confidence.

Read height optimization (Z height)

Z-height optimization automatically adjusts the detection height to maximize sensitivity and signal quality—while minimizing sample volume—ensuring reliable results even in low-volume or precious samples.

Microvolume & cuvette options

The SpectraDrop Micro-Volume Microplate for low-volume (2 to 4 μL) and SpectraCuvette™ Adapter Plate for cuvette-based applications eliminate the need for additional instrumentation.

Personalized touchscreen control

The intuitive touchscreen software simplifies tasks with Quick Read icons that save time when starting new experiments and a personalized library that stores frequently used protocols.

Western blot capable

The optional enhanced time-resolved fluorescence (TRF) detection module is optimal for quantitating as little as femtogram protein samples, providing improved sensitivity, extended signal stability and versatility to support a wide range of applications.

Absorbance

Common absorbance applications and assays including ELISAs, nucleic acid and protein quantitation, and microbial growth.

Cell Health

Cell Health

Cell viability refers to the number of healthy cells in a population and can be evaluated using assays that measure enzyme activity, cell membrane integrity, ATP production, and other indicators. These methods can employ luminescent, fluorescent, or colorimetric readouts as indicators of general cell viability or even specific cellular pathways. Cytotoxicity and cell viability assays are often used to assess a drug or other treatment’s effect, and are valuable tools in the search for new therapeutics, as well as advancing our understanding of how normal cells function.

Cell viability

Cell viability

Cell line development requires the discovery of single cell-derived clones that produce high and consistent levels of the target therapeutic protein. A critical first step in the process is the isolation of single, viable cells. Single cells proliferate to form colonies that can then be assessed for productivity of the target therapeutic protein. Viability and growth rates of single cell-derived clones can be characterized on the CloneSelect Imager and the SpectraMax i3x microplate reader and imaging cytometer.

Cellular Signaling

Cellular signaling allows cells to respond to their environment as well as to communicate with other cells. Proteins located on the cell surface can receive signals from the surroundings and transmit information into the cell via a series of protein interactions and biochemical reactions that comprise a signaling pathway. Multicellular organisms rely upon an extensive array of signaling pathways to coordinate the proper growth, regulation, and functioning of cells and tissues. If signaling between or within cells is dysregulated, inappropriate cellular responses may lead to cancer and other diseases.

DNA/RNA Quantitation

DNA/RNA Quantitation sample

The absorbance of a DNA sample measured at 260 nm on a spectrophotometer or microplate reader can be used to calculate its concentration. Absorbance quantitation of DNA works on samples ranging from about 0.25 µg/mL to about 125 µg/mL in a microplate format.

Drug Discovery & Development

Drug Discovery & Development Process

For every drug that makes it to the finish line, another nine don’t succeed. This alarming failure rate can be traced to reliance on 2D cell cultures that don’t closely mimic complex human biology, often leading to inaccurate predictions of a drug’s potential and extended drug development timelines.

ELISA

ELISA (Enzyme-linked immunosorbent assays)

Enzyme-linked immunosorbent assays (ELISAs) are used to measure the amount of a specific protein, using a microplate format, and results are most often detected via absorbance in the visible wavelength range. Chemiluminescent and fluorescent ELISA formats offer enhanced sensitivity for accurate quantitation of less abundant analytes.

Fluorescence

Many fluorescence-based assays include cell viability, GPCR activity, and fluorescent nucleic acid quantification.

Fluorescence Polarization (FP)

Fluorescence polarization (FP) is a technique that is widely used to monitor binding events in solution. It can be used to assess biomolecular interactions, including protein-antibody binding and DNA hybridization, as well as enzyme activity, and it has been adapted to basic research as well as high-throughput screening.

HTRF

HTRF®, an assay technology developed by Cisbio, combines standard FRET (fluorescence resonance energy transfer) with time-resolved measurement of fluorescence, eliminating short-lived background fluorescence. HTRF’s applications include assessment of small phosphorylated peptides, immunoassays for quantifying large glycoproteins, and indirect detection of tagged complexes such as CD28/CD86 via secondary antibodies. These homogeneous assays do not require wash steps and are amenable to small-volume microplate formats used in high-throughput screening.
(This assay is suitable for SpectraMax® iD5 and SpectraMax® Paradigm readers.)

IgG Quantification

Immunoglobulin G (IgG) Quantification

In therapeutic protein engineering and cell line development, IgG quantification is the process of measuring the amount of immunoglobulin G (IgG), a common class of therapeutic proteins, produced by a genetically modified cell line. This is important for evaluating and monitoring the productivity of the cell line and selecting the best candidate for further development of therapeutic antibodies.

Luminescence

The key luminescence-based assays include dual-luciferase reporter gene, chemiluminescent ELISA, cytotoxicity, and BRET.

Microbiology and Contaminant

Microbes, including bacteria, have been estimated to make up about 15 percent of the earth’s biomass, and microbes in the human body outnumber human cells by 10 to 1. These microorganisms provide great benefit to us and are also vital to many fields of research from medicine to alternative energy production. On the other hand, monitoring for microbes and the toxic substances they produce is necessary to ensure the safety of pharmaceutical products. Scientists whose research relies on mammalian cells must carefully monitor these cultures for unwanted microbial contaminants to ensure that their experimental results are reliable.

Nucleic Acid (DNA/RNA) Quantitation and Analysis

Nucleic acids are large biomolecules common to all known life forms. Deoxyribonucleic acid (DNA) consists of a double strand of pairs of nucleotides, while ribonucleic acid (RNA) is typically a single strand. In DNA, the nucleotides are adenine, cytosine, guanine, and thymine, while RNA contains uracil instead of thymine. DNA makes up the genetic material of all organisms, encoding the information cells need to synthesize proteins.

Protein Detection, Quantitation and Analysis

Protein detection, quantitation, and analysis are central to the investigation of a wide variety of biological processes. Measuring the concentration of protein is necessary to processes ranging from protein purification and labeling to sample preparation for electrophoresis. Protein can be quantitated directly via absorbance at 280 nm, or indirectly using colorimetric (BCA, Bradford, etc.) or fluorometric methods offering advantages such as greater sensitivity. To identify and measure a specific protein within a complex sample, for example, serum or cell lysate, an ELISA may be used.

Reporter Gene Assay

Reporter gene assays enable researchers to determine whether a gene is expressed and at what level. They are used to study signaling pathways, gene regulation, structure of regulatory elements, and much more. A reporter gene vector consists of a promoter or gene of interest, along with a reporter gene such as firefly luciferase, which codes for a protein whose activity can be easily measured. The vector is transfected into mammalian cells, and when the gene of interest is active in the cells, the reporter gene is also expressed and can be measured using appropriate detection reagents.

TRF, TR-FRET & HTRF

Time-resolved fluorescence – TRF and TR-FRET including HTRF assays provide applications resources to aid in your research, including kinase assays, cellular signaling pathways, protein-protein interactions, cell cytotoxicity, and more.

Western blotting

Western blotting is a popular technique used for protein detection and quantitation. Molecular Devices introduce a unique Western blot detection system on a multi-mode microplate reader, including colorimetric, fluorescence, and chemiluminescence detection methods.

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