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

Recent News

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Imaging neural development in the network

Fri, 10/23/2015

© 2015 Wiley Periodicals, Inc.

MADISON—A new study in the laboratory of Associate Professor Grace Boekhoff-Falk from the Department of Cell and Regenerative Biology at UW-Madison examines the expression of homeobox gene Distal-less II (DII) in the context of neural network development in the fruit fly Drosophila using confocal laser scanning microscopy (Department of Medical Microbiology Immunology) and live imaging by Jayne Squirrell with the Optical Workstation multiphoton microscope at LOCI.

Seeing pancreatic cancer in a new light

Wed, 09/16/2015

Cole Drifka and LOCI Co-Investigator W. John Kao use quantitative second harmonic generation microscopy to investigate the tumor microenvironment in pancreatic cancer. Image shown: front of a pancreatic ductal adenocarcinoma tumor infiltrating into the duodenal muscularis externa is characterized by distinct aligned and elongated collagen fibers orientated in the direction of invasion. Scale 100 μm. (Nature Publishing Group, 2015).

It's not the code, it's the community: 2015 ImageJ User and Development Conference

Fri, 09/04/2015

Over 200 attendees joined the 2015 ImageJ User and Development Conference held at the Wisconsin Institute for Discovery and sponsored by FEI, OlympusApplied Scientific Instrumentation, and Photometrics.  The conference was live tweeted by organizers and attendees and coorganized with the Morgridge Institute for Research, the University of Wisconsin-Madison, and the Luxembourg Institute of Science and Technology. See the interactive slideshow.

Navigating the collagen jungle

Tue, 09/01/2015

Image shown: End view and side view of laser propagation direction relative to collagen fiber axis. (Optics Letters 2015)Image shown: End view and side view of laser propagation direction relative to collagen fiber axis. (Optics Letters 2015)

MADISON--A new approach developed in the laboratory of LOCI collaborator Paul Campagnola uses reflective micro-prisms to discern collagen organization from multiple angles. 

Communing in the ImageJ ecosystem

Sun, 08/23/2015

Image Shown: Montage of collagen imaged by second harmonic generation of breast tumors in a mouse mammary model. Image is of 81 images stitched together using the maximum intensity projection feature and the stitching plugin of Fiji.Image Shown: Montage of collagen imaged by second harmonic generation of breast tumors in a mouse mammary model. Image is of 81 images stitched together using the maximum intensity projection feature and the stitching plugin of Fiji.

Metabolic imaging of cancer with Melissa Skala

Sat, 08/22/2015


Melissa Skala
, Assistant Professor of Biomedical Engineering at Vanderbilt will be coming to give a seminar next Tuesday August 25th at noon. Her seminar will be in the Tong Auditorium of Engineering Centers (Room 1003 Engineering Centers). 

Multiscale in-vivo optical imaging and microscopy with Elizabeth Hillman

Fri, 08/21/2015

Elizabeth Hillman, Associate Professor of Biomedical Engineering at Columbia will be coming to give a seminar next Friday August 28th at 2pm. Her seminar will be in the 3rd floor teaching lab of WID (please take elevators right next to Aldos coffee shop in WID). 

Innovating emergency medicine

Thu, 08/20/2015

MADISON (WKOW) -- Thanks to new funding at UW, doctors will be able to have some everyday wishes granted.  Engineers and students are working on prototypes for medical innovations that doctors have said they are lacking in their practice.  The UW Department of Emergency Medicine is teaming up with UW's Morgridge Advanced Fabrication Lab or "Fab Lab" to improve these medical tools, which could improve your time in the hospital.  

Illuminating mystery moon caves

Tue, 08/18/2015

MADISON—Andreas Velten, a Morgridge medical engineering affiliate and computational optics scientist with the University of Wisconsin-Madison Laboratory for Optical and Computational Instrumentation (LOCI), has developed a technology that fires and recaptures scattered laser light to literally "see around corners."

2015 ImageJ User and Developer Conference

Tue, 08/11/2015

MADISON—The 2015 ImageJ User and Developer Conference hosted by LOCI at the Wisconsin Institute for Discovery complements the EU meetings in Luxembourg and will offer workshops to improve software knowledge and usage covering both beginner/user and advanced/developer topics.

Upcoming Events

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Software at LOCI

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

SciJava Common
SLIM Curve
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:

Spectral Confocal
Optical Work Station

Browse all LOCI instrumentation

Research Highlight

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.

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.

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. 

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. 

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).

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.

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. 

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.

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