Light-guide Image Processing (LIP) 3D printed snapshot spectrometer for molecular imaging The overall goal of this research is to develop a new generation of a high performance, compact, snapshot hyperspectral imaging device that is compatible with research grade light microscopes. To accomplish this goal we will build a Light-guide 3D printed Imaging Processing spectrometer. The LIP Spectrometer will leverage two innovative technologies: fiber optics and 2-photon additive manufacturing to allow effective fabrication of custom fiber bundles providing different input vs. output organizations. These custom bundles will capture densely packed input and yield an arbitrary output with void spaces that allow spectral information to be spread out. Thus, the LIP device will convert an image from 2D image (input) to separated spatial-spectral information (after dispersion) on a large format sCMOS/CCDs image sensor. The proposed approach requires no significant computation or processing to create the (x, y, λ) data cube. Simple data re-organization will be sufficient to create spectral cubes. LIP spectrometer will be a widefield method acquiring full spectral information simultaneously from every pixel and therefore it will offer significant advantages in imaging speed and signal collection. The LIP spectrometer will allow changing spectral / spatial sampling through different fiber bundle and disperser configurations. The project will pursue two specific aims: (1) developing a proof-of-concept LIP spectrometer (250x250x60 over visible range 470—670nm) and (2) testing the LIP Spectrometer the dynamic range, imaging speed and sampling over in reference to other techniques like IMS or scanning methods. We will also perform imaging in several living cell imaging applications. These experiments will focus on tests in dynamic biological systems, specifically to study [ATP]/[ADP] ratio, cAMP signaling, and Ca2+ dynamics investigated in Dr. Piston’s lab. We will also quantitatively evaluate the results from the LIP, against the Zeiss LSM880, Optical Insights Spectral DV commercial systems and Image Mapping Spectrometry (IMS) developed previously by project’s PI (Dr.Tkaczyk).

The project targets the development of a high sensitivity, rapid acquisition imaging spectrometer enabling high resolution spectral imaging of biological samples. In consequence researchers will be able to advance the investigation of live cells with multiple fluorescent contrasts. The instrument’s principle allows obtaining spectral information for entire image without scanning and thus improves signal to noise ratio and allows more efficient investigation of transient biological events.