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AAT Bioquest

FluoroQuest™ Antifade Mounting Medium

FluoroQuest™ Antifade Mounting Medium is a specialized and stable formula that effectively preserves fluorescence by preventing rapid photobleaching of fluorescent proteins and dyes. This medium is characterized by an easy-to-use liquid form and readiness for immediate use without requiring warming. Additionally, it can be stored for long-term analysis without sealing and is compatible with simple sealing methods such as nail polish or plastic sealants. To ensure optimal performance, mounted slides should be stored at 4 °C and protected from light.

Example protocol

AT A GLANCE

Protocol Summary
  1. Prepare Samples (slides or microplate wells).

  2. Add a drop of a component and mount.

  3. Examine the specimen under microscope.

Important Note

These ready-to-use anti-fading reagents can be applied directly to the washed specimen. Although the reagents have been tested with lots of fixed samples, their optimal anti-fading efficiencies strongly depend on the properties of your samples. We suggest that you try more than one component for your imaging samples to get the ideal component. For example, one component may be more compatible with a fluorescently labeled antibody conjugate (or an enzyme substrate or special mounting specimens that contain lipophilic plasma membrane stains like DiI) than another one.

CELL PREPARATION

For guidelines on cell sample preparation, please visit:

https://www.aatbio.com/resources/guides/cell-sample-preparation.html

SAMPLE EXPERIMENTAL PROTOCOL

  1. Thaw the vial of FluoroQuest™ Antifade Mounting Medium at room temperature, protected from light.

  2. Remove any excess liquid from your specimen. Add a small drop of the selected component to the specimen. If the sample is on a slide or tissue culture dish, carefully place a coverslip on the drop, avoiding air bubbles. If the sample is on a coverslip, invert the coverslip on a clean glass slide. Remove any excess anti-fading component.

  3. The anti-fading reagents should be incubated for 1 hour to overnight. For long-term storage, seal the coverslip to the slide with nail polish or a plastic sealant. Mounted slides should be stored at 4°C in the dark for optimum sample longevity. The fluorescence imaging would remain stable for many weeks. Samples can be imaged immediately after mounting.

Page updated on November 23, 2024

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Catalog Number20007
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Storage, safety and handling

H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22

Storage

Refrigerated (2-8 °C); Minimize light exposure
Antifade properties were measured using cells stained with fluorescently labeled secondary antibodies, as listed in the graph. The antifade capability of the VectaShield and FluoroQuest™ Antifade mounting mediums was measured using a 20X objective with real-time imaging over 10 seconds of continuous exposure to the excitation illumination. The intensity after 10 seconds of exposure is expressed as 100% percent of the intensity at zero time. The data shown above in the graph represents the % fluorescence remaining after 3 minutes of continuous exposure.
Antifade properties were measured using cells stained with fluorescently labeled secondary antibodies, as listed in the graph. The antifade capability of the VectaShield and FluoroQuest™ Antifade mounting mediums was measured using a 20X objective with real-time imaging over 10 seconds of continuous exposure to the excitation illumination. The intensity after 10 seconds of exposure is expressed as 100% percent of the intensity at zero time. The data shown above in the graph represents the % fluorescence remaining after 3 minutes of continuous exposure.
Antifade properties were measured using cells stained with fluorescently labeled secondary antibodies, as listed in the graph. The antifade capability of the VectaShield and FluoroQuest™ Antifade mounting mediums was measured using a 20X objective with real-time imaging over 10 seconds of continuous exposure to the excitation illumination. The intensity after 10 seconds of exposure is expressed as 100% percent of the intensity at zero time. The data shown above in the graph represents the % fluorescence remaining after 3 minutes of continuous exposure.