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September/October 2011
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A fluorescence glass standard on an epifluorescent microscope.

Cell Signaling

Epifluorescence microscopes are used to generate fluorescence intensity images in several biology applications. A proposed new ASTM International standard will provide guidance on how to set up an experiment that uses an epifluorescence microscope to ensure that the imaging measurements are quantitative. The proposed standard will also provide guidance on how to use benchmarks to allow comparisons of measurements that are taken on different days with the same or similar microscopes.

The proposed new standard, WK33434, Guide for Performing Quantitative Fluorescence Intensity Measurements in Cell-Based Assays with Epifluorescence Microscopy, is under the jurisdiction of Subcommittee F04.46 on Cell Signaling, part of ASTM International Committee F04 on Medical and Surgical Materials and Devices.

“Fluorescence imaging with epifluorescence microscopy is routinely used to quantify relative concentrations and the spatial location of molecules and protein in biological samples such as cells,” says John Elliott, research scientist, biochemical science division, National Institute of Standards and Technology, and a co-chairman of F04.46. “To use this technique reliably, it is critical that the optical and imaging components of the system are sufficient to generate quantitative fluorescence images. We initiated WK33434 to provide important guidance tips for ensuring fluorescence imaging measurements of biological samples are quantitative and can be compared to one another.”

Elliott notes that F04.46 actively seeks participation in its standards development activity.

“We are interested in participation from microscopy instrument manufacturers, fluorescence microscopy users, image analysis software developers, cell biologists, material scientists and scientists involved in standards development for fluorescence microscopy,” says Elliott.

CONTACT

Technical Information: John Elliott, National Institute of Standards and Technology, Gaithersburg, Md.

Phone: 301-975-8551

ASTM Staff: Pat Picariello

Phone: 610-832-9720