Multiplex imaging antibody technology

Stain. Image. Release. Repeat.
Antibody complexes with releasable fluorochromes
For cyclic immunofluorescence staining using MICS technology

Why use REAdye_lease™ Releasable Fluorochromes for cyclic immunofluorescence?

• Allow signal erasure from an entire stained sample in a single step
• Functionally validated for MICS on FFPE, PFA-, or acetone-fixed samples
• Lot-consistent and reproducible due to recombinantly engineered antibody fragments

How it works: The basic principle

REAdye_lease Releasable Fluorochromes are fluorochrome-conjugated antibody complexes designed for MACSima™ Imaging Cyclic Staining (MICS), our cyclic immunofluorescence technology, using the MACSima Imaging Platform.

They consist of recombinantly engineered antibody fragments that are coupled to releasable fluorochromes, together forming the REAdye_lease Antibody Complex.

All REAdye_lease Antibody Complexes are pre-validated on either formalin-fixed-paraffin-embedded (FFPE, IHC-p), PFA-fixed, or acetone-fixed (IHC-f or ICC) samples.

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After fixed samples are positioned on MACSwell Sample Carriers, they are ready to be processed by our fully automated MACSima Imaging System.
In the first staining cycle, an initial set of markers is stained with REAdye_lease Antibody Complexes. (For simplicity, a one-color staining against a single marker is depicted here; However, multiplexing of several markers per cycle is possible.)

In the next step, an image of the stained sample is acquired.

After image acquisition, the fluorescent signal produced by the REAdye_lease Antibody Complexes is erased by controlled release of fluorochromes from the antibody fragments. This is achieved by addition of the REAlease® Release Reagent, which triggers gentle release of fluorochromes from the entire sample in a single step. This very mild treatment does not affect sample or epitope integrity. After fluorochrome release, several washing steps ensure that all fluorochromes are removed and that the detection channels are free for the next staining cycle. The MACSima Imaging System will automatically continue with all following programmed cycles, thereby acquiring hundreds of marker images for one sample.

REAdye_lease Antibody Complexes are validated for our MICS protocol, which is based on a quick, 10-minute immunolabeling immunostaining step at room temperature, and are therefore also perfectly suited for standard immunofluorescence applications.

Performance data for REAdye_lease Releasable Fluorochromes

Uncompromised fluorescent antibody staining and efficient signal release from target cells

REAdye_lease Antibody Complexes provide high signal-to-background ratios and minimal non-specific binding. Incubation of the sample with the REAlease Release Reagent for 10 min results in a quick and highly efficient fluorochrome release from the entire sample at once, ensuring efficient signal erasure and a free detection channel ready for use in the next staining cycle.

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