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Metabolic FLIM of Macroscopic Objects

Fluorescence lifetime images of macroscopic samples as large as 20 mm can by the DCS‑120 MACRO FLIM system. The system can be used for measuring metabolic parameters of biological tissue. Metabolic FLIM is performed via the fluorescence decay parameters of NAD(P)H and thus requires an excitation wavelength of 375 nm or shorter. Commonly used lens elements have poor transmission and large chromatic aberration at this wavelength. We therefore replaced the standard scan lens with one that consists of a combination of UV achromats. This increases the efficiency of the system by 40%. The spatial resolution is about 15 µm, images can be recorded with pixel numbers as large as 2048 x 2048. Acquisition times range from a few seconds for imaging with low pixel number and moderate lifetime accuracy to a few minutes for mega-pixel images and ultra-high accuracy of the decay data. We demonstrate the use of the system for metabolic imaging of a tumor in a mouse and of glioblastoma in a rat brain.

Keywords: FLIM, Label-Free, NADH, Macroscopic Objects, Tumor, Glioblastoma

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