Welcome to the Laboratory for Optical and Computational Instrumentation (LOCI) web site! We are a biophotonics instrumentation laboratory, aiming to develop advanced optical and computational techniques for imaging and experimentally manipulating living specimens.
The purpose of this standalone MATLAB package is to allow users to automatically extract collagen fibers in an image, and quantify fibers with descriptive statistics, such as fiber angle, fiber length, fiber straightness, and fiber width.
The Data Browser is an ImageJ plugin that facilitates quick browsing of multichannel, multi-focal plane time course datasets. It is integrated with the Bio-Formats importer plugin—as well as ImageJ's built-in hyperstack and virtual stack support—to provide multidimensional visualization capabilities across space, time and channels.
Fiji is an image processing package. It can be described as a distribution of ImageJ (and ImageJ2) together with Java, Java3D and a lot of plugins organized into a coherent menu structure. Fiji compares to ImageJ as Ubuntu compares to Linux.
ImageJ2 is a new version of ImageJ for the next generation of multidimensional image data. It improves upon ImageJ's core design, enabling support for N-dimensional image data beyond 5D, from sources beyond just hard disks, additional pixel types, more flexible visualization, more modular analysis, and headless capabilities for automated server-side image processing.
The Intensity Macro for Background Subtraction is customized for the image processing and analysis done by the Ogle Lab. It prompts the user to choose a background region of interest, a percentage of background to retain and a region of interest to which the subtraction should be applied. The intensity level to subtract from the fluorescent regions of interest is calculated by computing the level at which the background intensity histogram corresponds to the amount of background the user desires to retain.
SCientific Image Formats Input and Output (SCIFIO) is a framework for developing and accessing image I/O plug-ins. SCIFIO will include support for many open-source formats, and Bio-Formats will become the flagship SCIFIO plug-in.
The Threshold and Calculate Average Intensity with Brush Tools macro is customized for immunohistochemistry image analysis done in the Ogle Lab. It is especially suited for stained tissue sections that have large areas of background interspersed between areas of signal.
This script creates an animation from ROIs added to the ROI Manager. It uses the associated image plane of each ROI, adjusted spatially such that the first (X, Y) coordinate of each ROI occupies the same location, as a "poor man's" registration technique. Each plane is then assembled into the final movie.
The TumorTrace program is an automated image analysis tool, developed in MATLAB (The Mathworks, Inc., Natick, MA), for examining the ECM surrounding cells or tumors in the context of cellular morphology, protein expression and movement. It takes as input multiple image channels, either single images or stacks representing time-series or 3D data. It then finds a metric for the cell/cell cluster morphology and outputs plots representing intensity, morphology, collagen fiber alignment, and cell movement; .csv files containing raw data and image files containing the regions of interest.
As part of the development of ImageJ2, we are reworking VisBio
as a collection of ImageJ plugins. There is an initial version of one
such plugin, VisBio Ortho Stack, available as part of the LOCI Fiji plugins.
Bioimage informatics is an interdisciplinary field of research encompassing biology, information science, computer science, statistics, and engineering. Bioimage informatics strives to automate, simplify, and otherwise improve and reinvent techniques for the description, management, analysis, and preservation of biological image data.
Eukaryotic cells depend upon the mitochondrial electron transport chain to produce energy in the form of ATP when oxygen is present. The first complex of this chain oxidizes NADH to NAD+. Conveniently, NADH is an intrinsically fluorescent molecule, while NAD+ is not. Time-resolved studies of NADH fluorescence using fluorescence lifetime imaging (FLIM) can be used to obtain information about NADH and metabolic states in non-malignant and malignant cells.
Collagen organization and density can have a profound effect on the behavior of breast cancer cells. Using second harmonic generation (SHG), the structure of collagen in intact mammary mouse mammary glands can be visualized. In collaboration with the Keely Lab, a metatstatic collagen signature (TACS3) has been identified leading to useful characterization of breast tissue and tumors.
Several research projects currently being pursued at LOCI require fast, multiphoton imaging. On the basis of the rationale outlined below we are currently developing a high-speed, single beam laser-scanning MP system. The scanning system will be used in conjunction with the spectral/lifetime detector and will be tightly integrated with this device.
Traditional flow cytometers
are unable to accommodate cell aggregates and often require extrinsic
fluorescent labeling for data analysis.
In an effort to analyze larger cell aggregates, a multiphoton flow cytometry
(MPFC) instrument has been constructed.
The system is comprised of a flow cell through which large particles and
aggregates travel, an optics system with multiphoton excitation capabilities,
and data acquisition software. The
flow cell is mounted on an adjustable stage insert compatible with most
While diagnosis of human breast cancers is advancing, current biomarkers cannot predict outcome for all patients. Endogenous optical properties of cells and tissues could potentially be used as biomarkers in the clinic. Collagen can be detected, without staining or labeling of tissue, using second harmonic generation (SHG) imaging, a multiphoton technique. Collagen signatures, see in mouse models, could be used in breast cancer patients to predict outcomes.
Spectral imaging is the collection and display of the spectral components of a fluorescence image. LOCI is currently developing a combined spectral/lifetime detector that is optimized for low-light level multiphoton imaging. The detector works in photon counting mode and essentially sorts detected photons into spectral and temporal bins. This detector is being developed primarily for the Optical Workstation but will also be used with the high-speed multiphoton imaging system currently under development.
In 1994, we commissioned a multiphoton imaging system that featured an all solid-state excitation source, a 1047nm Nd:YLF laser. The laser was developed to our specifications by Prof. Allister Ferguson's group at the University of Strathclyde, Scotland, and by Microlase Ltd., a company founded by Prof. Ferguson. This was the first all-solid-state multiphoton system to be developed (Wokosin et al., 1996. Proc.SPIE 2678, 38).