Fluorescence Imaging MicroSpectrophotometer (FIMS)

FIMS is based on an epifluorescence microscope and utilizes a cooled CCD camera, computer interfaced monochromator, and a Circular Variable Interference Filter (CVIF). Windows based software controls data processing and display such that the fluorescence excitation and / or fluorescence emission spectrum of every pixel in a field of view can be determined simultaneously. This software also includes tools for extracting features and sorting spectra by various criteria which can be visualized through the use of color contour plots. Preliminary data have been acquired with an early prototype of FIMS on diverse samples, including fluorescent microspheres (see figure, below) and bacteria expressing fluorescent proteins. An HTML copy of this manuscript, which is readable by almost all browsers, can be found on this web site, and a reprint of a published manuscript is available to browsers which support MS Word or PDF.

The figure shows a FIMS analysis of a mixture of 6 different fluorescent beads. Panel A uses hyperspectral pseudocoloring techniques to display each category of beads as a different color. This color code is consistent with the vertical bar to the right of the contour plot in Panel B, which shows the (desampled) spectral emission intensity of single pixels as a function of wavelength. The color coding is also maintained in Panel C, in which the average spectrum for each of the 6 types of beads plus background (pseudocolored black) is plotted.

FIMS patents are issued and pending.

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