Authors: Vince Rossi, Spencer Brown
Mentors: Vince Rossi
Insitution: Utah Valley University
Current systems of generating 3D fluorescent images of samples require scanning of a sample or multiple exposures to acquire a series of images. Collecting a 3D fluorescent image in these fashions is timely, ranging from minutes to tens of minutes. This is not ideal for imaging live, dynamic samples that change as time progresses. The use of holographic techniques allows for the collection of a 3D image on the scale of milliseconds, therefore negating motion artifacts. Combining the speed of digital holography with fluorescent imaging will allow for identification of labeled biological components within a sample as well as the ability to view the sample in a 3D space thus offering greater detail and information on the sample. This process is called Holographically Informed Fluorescence Imaging (HIFI). Simulations have been run using MatLab in which a sample is imaged using Phase Shifting Digital Holography. The 3D reconstruction of the image is propagated using Huygens Convolution Method, and then all of the pixels of the image are brought into focus numerically. This image is then overlayed with the fluorescence image obtained. Each pixel of the fluorescent image is propagated to the depth of focus for each pixel within the 3D holographic image and a 3D fluorescent image is obtained. Simulations of this process have proved successful. A proof of principle experiment is currently underway as to prove the effectiveness of this fluorescent imaging method.