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Fluorescent Polarization

Fluorescent Polarization

Sanford Burnham Medical Research Institute’s Fluorescent Polarization in Microgravity (NanoRacks-Micro-gRx) research examines microgravity’s effect on fluorescent polarization, which paves the way for advanced biology research and drug development in space.

Customer: Sanford Burnham Research Institute
Research: Fluorescent Polarization
NanoRacks Facility: NanoRacks Plate Reader
Mission Duration: 09/2014 – 03/2015
Mission Status: Complete
More Info: From NASA website


Research Overview

The research validates the NanoRacks Plate Reader facility in three of its five modes of operation in order to examine the effect of microgravity on fluorescent polarization (FP) for a fluorophore in solution, to validate the UV-Visible mode by using three different absorbance wavelengths in serially diluted liquids, and to validate the fluorescence intensity by using serial dilutions of two different fluorophores.

This investigation also serves as part of the process of validating and establishing a workflow for the NanoRacks Plate Reader facility. Testing is conducted with four microplate samples.

This helps assess if molecular processes are the same in space as on Earth. This opens the door for future advanced biology and pharmacology research in microgravity. By transferring advanced technologies to the International Space Station (ISS), researchers are able to determine the effectiveness of medicines in microgravity and explore biochemical and cellular pathways that can be targeted for new disease therapies.

Description
NanoRacks-Fluorescent Polarization in Microgravity (NanoRacks-Micro-gRx) experiments validate at least three of the five operating modes for the Molecular Devices Spectramax M5 multimode microtiter plate reader (NanoRacks Plate Reader), quantify the association of biological molecules as detected by fluorescent polarization (FP), and test the functionality of a 384-well microplate. These experiments are described by the following three aims:

  1. Perform a fluorescent polarization experiment which validates the FP mode of the plate reader and provide experimental comparison for detecting the binding of molecules by FP in microgravity. FP measurement of a fluorescently labeled ligand binding to a larger molecule provides information on molecular orientation and mobility in solution of the bound complex versus the free ligand. An exact replica of the experiment is tested at the same time point on earth using the same instrument to detect changes in FP due to microgravity.
  2. Validate the UV-Visible mode by using three different absorbance wavelengths in serially diluted liquids. Three absorbance wavelengths, which are the most commonly used by scientists, are tested.
  3. Validate the fluorescence intensity mode by using serial dilutions of two different fluorophores (fluorescent chemical compounds that re-emit light upon light excitation). The two most commonly used fluorophores are tested during this portion of the M5’s validation.

These experiments are necessary to validate that the NanoRacks Plate Reader is operating correctly and to its specifications. The results generated from these experiments have interest to physicists, biophysicists, biologists as well as scientists working in the field of drug discovery who wish to use the plate reader for future life science experiments.

For the testing of the fluorescence polarization module of the NanoRacks Plate Reader, the use of a fluorescently labeled biotin conjugate and an antibody to biotin are employed. Biotin also known as vitamin H, coenzyme R or vitamin B7 is involved in the synthesis of fatty acids and glucose. This modified version of this molecule labeled with the fluorescent dye fluorescein is commercially available.

Applications
The ways in which certain molecules change in response to light can tell scientists something about their orientation, movement and interactions with other molecules. Scientists use this technique, called fluorescence anisotropy, to study new drugs to treat disease. But many molecules behave differently in space than they do on Earth.

This investigation studies how microgravity affects molecules and the process of fluorescence anisotropy. Understanding differences between this technique in microgravity and on Earth will help researchers study new medicines and explore new cell therapies in space.

Molecules and cells may behave differently in space, allowing researchers to perform unique experiments and to study biology in new ways. Results from this investigation open the door for future advanced research in biology and pharmacology, or the study of drugs, in microgravity.

Developing new drugs in space may lead to new treatments for a wide range of human diseases, benefiting people on Earth.

Read more about the Fluorescent Polarization Research at the NASA website.

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