🏆 Foundational Paper

An interactive visualization tool for multi-channel confocal microscopy data in neurobiology research.

Wan Yong, Otsuna Hideo, Chien Chi-Bin, Hansen Charles

📰 IEEE transactions on visualization and computer graphics 📅 2009 📊 101 citations

Abstract

Confocal microscopy is widely used in neurobiology for studying the three-dimensional structure of the nervous system. Confocal image data are often multi-channel, with each channel resulting from a different fluorescent dye or fluorescent protein; one channel may have dense data, while another has sparse; and there are often structures at several spatial scales: subneuronal domains, neurons, and large groups of neurons (brain regions). Even qualitative analysis can therefore require visualization using techniques and parameters fine-tuned to a particular dataset. Despite the plethora of volume rendering techniques that have been available for many years, the techniques standardly used in neurobiological research are somewhat rudimentary, such as looking at image slices or maximal intensity projections. Thus there is a real demand from neurobiologists, and biologists in general, for a flexible visualization tool that allows interactive visualization of multi-channel confocal data, with rapid fine-tuning of parameters to reveal the three-dimensional relationships of structures of interest. Together with neurobiologists, we have designed such a tool, choosing visualization methods to suit the characteristics of confocal data and a typical biologist's workflow. We use interactive volume rendering with intuitive settings for multidimensional transfer functions, multiple render modes and multi-views for multi-channel volume data, and embedding of polygon data into volume data for rendering and editing. As an example, we apply this tool to visualize confocal microscopy datasets of the developing zebrafish visual system.

🔬 Techniques

✨ Fluorophores

GFP

🧪 Sample Preparation

💻 Software Details

Image Analysis:
Imaris Amira

🏛️ Research Organizations (ROR)

Affiliated research institutions:

📊 Figures

Fig. 1

A typical workflow in neurobiology research with confocal microscopy.

Fig 2

Rendering effects. A: Default shading effect; B: Lowering the ambient intensity increases the contrast for local features; C: Depth cueing can better show the overall shape; D: Cyan colored channel (a...

Fig. 3

2D Transfer function and its parameters.

Fig. 4

Render modes. A: Layered; B: Depth sorted; C: Composite. In layered mode (A), the rendering order of the channels, from top to bottom, is: neurons (green), muscles (red), and all cell nuclei (blue). T...

Fig. 5

Depth peeling algorithm. A: Only one depth peeling layer; B: n depth peeling layers.

Fig. 6

Depth peeling results. A: Ventral view of the volume data showing retinal ganglion cells connecting between the eye and the brain; B: Polygon data added, separating volume data into eye (magenta) and ...

Fig. 7

Volume culling with polygon data. A: Original volume data; B: Volume data after culling; C: The process of culling the occluding data in the yellow channel, showing retinal ganglion cells (RGCs; C1: O...

Fig. 8

Different transfer function settings of volume data in different regions. The process shows how cells in different biologically meaningful regions are marked out, where the color-coded volume data rep...

Fig. 9

A screenshot of our tool. A: Toolbar, and the buttons are: Open Volume, Open Project, Save Project, Open Mesh, New View, Make Movie, and Info. B: Data View, loaded datasets are listed. C: Scene View, ...

Fig. 10

The zebrafish visual system, rendered with our tool (Au2013C) and compared to previously used maximum intensity projections (Du2013F). A, D: Dorsal views of neurons expressing Tg(brn3a-hsp70:GFP) (gre...

Fig. 11

The rendering result comparison between commercial packages and our tool. Left column: renderings of three channels (red: muscles, green: neurons, blue: all cell nuclei). The dashed rectangular region...

Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.

🏛️ Imaging Facility

🏛️ University of Utah

💬 Discussion

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