Welcome to the Laboratory for Optical and Computational Instrumentation!
We are a biophotonics instrumentation laboratory developing advanced optical and computational techniques for imaging and manipulating living specimens.

Recent News

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McPherson Eye Research Institute Endowed Lecture: Talking to the brain in its own language with Sheila Nirenberg

Mon, 05/18/2015

Understanding how electrical pulses are generated in neural coding provides insight in prosthetic device development for neuroscientist Sheila Nirenberg.


Advancing imaging technology

Wed, 05/06/2015

“We have an imaging system and a lighting system paired with it,” says Morgridge research associate Adam Uselmann. “It allows you to have a well-lit room while you’re doing fluorescence imaging, something that normally would have to be done in the dark.”

Probing nanoscale tissue structure using light scattering

Tue, 04/28/2015

 

















Steven L. Jacques will present his work using light scattering to illuminate understanding of size distribution and granularity of structures in the tissue.

Developing faster multipoint scanners for live cell imaging

Mon, 04/20/2015

Acquisition of confocal images of time-based volumetric data can provide a more complete picture of cellular events than imaging in a single focal plane, even with monolayers of cell cultures.

Visualizing tumor metabolism

Wed, 04/15/2015

 

 

 

 











Many of the signaling pathways that promote aggressive behavior in cancer cells also regulate metabolism in the cell.  

Monitoring multiscale changes in the cervix

Mon, 04/13/2015








Researchers in the laboratory of Medical Physics Professor Timothy Hall use second harmonic generation (SHG) images registered with quantitative ultrasound (QUS) data to comprehensively characterize cervical microstructure in high resolution.

The Why Files announces 2015 Cool Science Image Contest Winners

Fri, 04/10/2015


Scientific imagery is a critical form of data in many fields and can yield striking insights into nature and the way things work.

2015 Midwestern Microscopy and Microanalysis Meeting

Thu, 04/09/2015



LOCI Director Kevin Eliceiri and LOCI Co-Investigator John White were featured speakers in this year's Midwest Microscopy and Microanalysis Meeting on multiphoton imaging. M3S March 2015 Newsletter

Ellen Arena named Morgridge Institute for Research postdoctoral fellow

Tue, 04/07/2015

Ellen Arena begins her new chapter at Morgridge, focusing on microscopy and bioimaging of viral-host cell invasion. She will work directly with Kevin Eliceiri and Ming Yuan of the Morgridge Institute; and with Nathan Sherer, a UW-Madison assistant professor of oncology.

Software at LOCI

LOCI is an active partner in the SciJava and Open Microscopy Environment consortiums, and participates in many related software projects, including:

ImageJ
Fiji
SCIFIO
Bio-Formats
WiscScan
ImgLib2
SciJava Common
SLIM Curve
KNIME
OMERO
Insight Toolkit

Browse all LOCI software

Software Highlight

Bio-Formats is a library—and collection of ImageJ plugins—for reading and writing popular microscopy file formats, including processing and conversion of metadata into standard OME-XML structures.

When do I use CurveAlign and when  do I use CT-FIRE?

These two programs were developed with complementary but slightly different main goals. CurveAlign was developed first and had the main goal of quantifying all fiber angles within a region of interest relative to a user defined boundary be it a straight line or a tumor boundary. As our research grew in investigating the role of collagen in cancer progression and invasion we wanted to investigate how individual fiber parameters could influence cancer and other diseases. Out of this need came the development of CT-FIRE to analyze individual fiber metrics such as length, width, angle, and curvature.  Besides the relative angle quantification, the newest version of CurveAlign can be used to extract other collagen fiber features, such as localized fiber density, fiber alignment, and the spatial relationship between fiber and the defined boundary. In addition, the extracted individual fibers extracted by CT-FIRE can be imported into the CurveAlign for the feature extraction mentioned above. We have future plans to integrate these programs further. For now CurveAlign should be used for bulk assessment of collagen features including angles/density and CT-FIRE for individual fiber quantification.

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.

The image stitching plugins, written by Stephan Preibisch, provide a means for reconstructing large mosaics from tiled datasets.

The LOCI Fiji plugins are a collection of plugins for Fiji, available from our Fiji update site.

The Fusion Event Locator & Classifier is an ImageJ macro & MATLAB function to identify fusion events in image stack & classify as proliferating or unchanging.

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.

Jar2Lib is a command line tool for generating C++ wrapper libraries around Java JAR files. We use it to generate the BF-CPP bindings for Bio-Formats, which we use in our WiscScan acquisition software.

A script to help manage Prairie Technologies datasets. It converts Prairie datasets to compressed OME-TIFF, and archives the original files as a ZIP archive (which can then be backed up externally).

SCientific Image Format 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.

SLIM Curve is an exponential curve fitting library used for Fluorescent Lifetime Imaging (FLIM) and Spectral Lifetime Imaging (SLIM).

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.

VisBio is a biological visualization tool designed for easy visualization and analysis of multidimensional image data.

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.

WiscScan is acquisition software for laser scanning microscopy, used by LOCI and our collaborators for much of our scientific research.

WiscScan Flow Cytometry is a standalone tool for flow cytometry analysis. It works as a plugin for ImageJ that enables post-acquisition analysis of flow cytometry data.

Instrumentation at LOCI

To benefit the scientific community in an accessible way, LOCI develops new and improved imaging instrumentation and optical-based experimental techniques:

SETI
OpenSPIM
CAMM
MPFC
Spectral Confocal
FLIM
Intravital
Photoactivation
Ultra
SLM
Optical Work Station
Beam-Shaping

Browse all LOCI instrumentation

Research Highlight

High Speed Multiphoton Imaging

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.

Spectral and Lifetime Imaging

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.

Metabolic Mapping

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 and Breast Cancer

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. 

Collagen Signatures

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.

Multiphoton Imaging System

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).

Multiphoton Flow Cytometry

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 microscope stages. 

Image Informatics

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.

National Center for Research Resources
National Institute of Biomedical Imaging and Bioengineering
National Institute of Health
National Science Foundation
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