Mabtech IRIS 2

Mabtech IRIS 2

Mabtech IRIS™ 2 – Spot analysis reinvented

IRIS 2 is our FluoroSpot and ELISpot reader capable of analyzing up to four analytes simultaneously with unparalleled accuracy and speed. With the new FociSpot application, you can now quantify virus foci in focus forming assays (FFA). Mabtech IRIS 2 utilizes our patented RAWspot™ algorithm, ensuring accurate spot center detection and thus, correct counting. Together with the intuitive software Mabtech Apex™, IRIS 2 provides a plug-and-play system that simplifies your workflow. Just load the plate, select the assay, and press "read". 

  • ELISpot, FluoroSpot (up to 4 colors), and FociSpot (FFA with enzymatic labeling)
  • Precise spot counting 
  • Fixed focus and self-calibrating XY table for a streamlined workflow
  • Intuitive software for a smooth user experience
  • Customizable presets for recurrent experiments
  • Ready for automated plate-loading

 

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Product specifications

Intended use

Serum/Plasma samples

Recommendation

Application 

ELISpot1 color
FluoroSpot

4 colors

Recommended fluorophores (excitation/emission):
380 nm/430 nm (DAPI equivalent)
490 nm/510 nm (FITC equivalent)  
550 nm/570 nm (Cy3 equivalent)
640 nm/660 nm (Cy5 equivalent)

FociSpot1 color  

 

 

Hardware 

XY tableSelf-calibrating
Light source(s)

FluoroSpot: LED380, LED490, LED550, LED640

ELISpot: White LED ring light

Camera sensorCMOS sensor with global shutter
Camera lens    Telecentric
Resolution (HxW)2048 x 2048 pixels
Plate types: 96-well 

ELISpot: MSIP, MAIPSWU

FluoroSpot: IPFL

FociSpot: Corning, Nunc

Dimensions (HxWxD)505 x 430 x 480 mm
ComputerDesktop PC included
Robotic automationReady

 

 

Software

Mabtech Apex™RAWspot™ technology included
Export formats.raw .jpg .xlsx .pzfx
Reading speed

ELISpot:
<2 min/plate

FluoroSpot:
1 color <5 min/plate
2 colors <7 min/plate
3 colors<10 min/plate
4 colors<13 min/plate

FociSpot:
<2 min/plate

Operating systemWindows

 

 

Service

Warranty

1 year, option to prolong

Qualification

IQ OQ PQ

Regulation

Compliance with CE, RoHS, REACH, WEEE, FCC, ICES, CFR21 part 11, EU Annex 11

 

Shipping and Storage

Features

 

Streamlined workflow

Mabtech IRIS 2 is pre-calibrated for a smooth workflow. Simply insert the plate, select assay type, and press the "read" button. The entire signal from every spots is recorded so that adjustments can be made after plate reading – you never have to re-read a plate. In addition, the Mabtech Apex™ software offers intuitive layout features and automatic data export to speed up the time from data acquisition to analysis. 

 

 

 

Precise spot counting  

The software for Mabtech IRIS 2 is based on our patented RAWspot™ technology. The RAWspot algorithm is optimized to find the exact location of the spot center. This refined spot center detection is the basis for accurate spot counting and is essential to identify cells secreting multiple analytes in FluoroSpot. 
Mabtech IRIS 2 also delivers a new dimension of spot data – the relative spot volume (RSV) – as RAWspot extracts a 3-dimensional volume for each spot corresponding to the amount of secreted analyte.

 

 

 

Read now, analyze later

After plate reading, you are free to adjust parameters such as brightness, contrast, spot intensity, and spot size thresholds without affecting the original data. As IRIS 2 has already recorded the uncompressed spot signals straight from the camera sensor, changes can be made later without affecting the original data. The fast and responsive Apex software has multiple file-save formats available simplifying data handling.

We take quality control seriously at Mabtech. Read about the different regulatory requirements (such as CFR21 part 11, EU Annex 11) Mabtech IRIS 2 meets here.

 

 

 

Minimized variability

Automated reader configuration and optimized presets minimize user-defined input and ensure an objective analysis independent of the user’s level of experience. Plus, the intuitive software facilitates smooth onboarding. Spot analysis has never been easier. 

 

 

 

Ready for automation

Mabtech IRIS 2 was designed to make the analysis of larger projects as painless as possible. In addition to self-calibration, responsive software, and easy data handling, IRIS 2 allows for robotic plate loading. Our reader can be part of an automated workflow with a robotic arm or be fully integrated into a larger automated setup. Each automation solution is unique, and we’ll be here to help every step of the way.

 

 

Performance qualification  

Our readers are reliable tools that can be trusted to deliver accurate results over an extended period of use. However, it’s essential to demonstrate that the readers maintain acceptable performance over time, especially when facing internal or external review for regulatory compliance. Mabtech’s Performance Qualification (PQ) plate provides a solution for verifying that our instruments fulfill quality specifications and to ensure reproducibility. The PQ plate allows users to conduct a series of quantitative and qualitative tests, making it easy to identify any incorrect or inconsistent operation of the reader. By using PQ plates, our instruments can be confidently validated, providing peace of mind knowing that experiments are accurate and reliable.

Different sections of the PQ-plate for IRIS:
1.    Tests focus and RAWspot algorithm in FluoroSpot
2.    Tests LED light intensity in FluoroSpot
3.    ELISpot test

 

 

Count foci with FociSpot

FociSpot is our new assay platform that includes both kits and a new analysis capability of Mabtech IRIS 2, allowing for automated counting of foci in a 96-well plate format. In FociSpot, foci are detected with immunostaining using virus-specific mAbs and a precipitating substrate reaction similar to ELISpot.

 

 

Find out which analyte combinations we have evaluated in T cell FluoroSpot and which combinations are affected by capture effects or not. 

What is a capture effect?

When capture antibodies with different specificities are coated together, the capture of one cytokine may affect the secretion of other cytokines. This is usually more pronounced when studying T cell responses with polyclonal stimuli compared to antigen-specific responses. Capture effects are seldom a problem and can often be counteracted by the addition of an anti-CD28 antibody.

Depiction of capture effects observed in FluoroSpot

(1) IL-2 secreted by the activated T cell is captured by coated anti-IL-2 capture antibodies. (2) As a result, IL-2-stimulation of the T cell itself (autocrine stimulation) as well as nearby T cells (paracrine stimulation) is impaired, ultimately leading to (3) decreased IFN-γ secretion.

Co-stimulation with anti-CD28

Anti-CD28 mAb provides a co-stimulatory signal to antigen-specific responses by binding to CD28 on T cells. The addition of an anti-CD28 mAb to the cell culture enhances antigen-specific responses and can counteract capture effects. For example, the presence of IL-2 capture antibodies may result in reduced activation of T cells, as capturing of IL-2 decreases the amount of available IL-2 and thereby dampens the IL-2-mediated stimulation of T cells. The addition of anti-CD28 mAb restores IFN-γ responses (depicted in the below images). Further optimization may be necessary, depending on the cells and stimuli used. Too high concentrations of the anti-CD28 mAb may result in an elevation of non-specific cytokine secretion.

Overcoming capture effects using anti-CD28 anitbodies in FluoroSpot

(1) An anti-CD28 antibody can be added to provide a co-stimulatory signal that can restore (2) e.g. IFN-γ responses.

How to investigate capture effects

Our FluoroSpot Plus kits are evaluated for capture effects, and in studies of T cell responses we recommend co-stimulation with anti-CD28. With FluoroSpot Flex, it is possible to combine and build your own kit. For guidance look at our analyte combination table (above). Capture effects can be investigated by quantifying spot numbers in wells coated with single capture antibodies and compared to wells coated with a mixture of the capture antibodies. The compensatory effect of the anti-CD28 mAb may be assessed by comparing cells cultured with and without the anti-CD28 mAb.

Tutorial, Published December 28, 2023

Incubation strategies for success in ELISpot and FluoroSpot
We've made a handy table summarizing incubation times for different analytes to make your research planning for the ELISpot and FluoroSpot assay a little easier.
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