Transcript
Confocal Application Notes Vol. 4
August 2006
New Viewer Tools in LAS AF
(From version 1.5.0 Build 767 and later) Prepared by Myriam Gastard, Ph.D. Exton Support Group, Leica Microsystems, Inc.
New tools are now available in LASAF. They will give you more freedom in viewing your images as well as more tools to review and analyze your data.
• Orthogonal Sectioning This tool will allow you to view your xyz series with the corresponding projections in XY, XZ, and ZY. Then any point can be chosen and will be projected in the corresponding projection in the other plans.
Viewer
Orthogonal Sectioning
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The “Orthogonal Sectioning” tool (1) is accessible from the Viewer window (right screen). This tool won’t appear without a series being selected in the experiment window. On the other hand, the orthogonal tool can be use on a previously saved series. It doesn’t need to be live data.
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Once activated the series are displayed in the different orthogonal plans. If the orthogonal 2projection channel is clicked twice on it (2), the projection will be displayed full screen.
Leica Microsystems Inc. 410 Eagleview Blvd, Ste. 107 Exton, PA 19341
Telephone (610)-321-0460 Toll Free 866-830-0735 Fax (610)-321-0425 www.confocal-microscopy.com
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Confocal Application Notes Vol. 4
August 2006
3The crosshairs (3) may be used to identify an object of interest and instantaneously find the same object in the different other orthogonal projections.
XY
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ZY
XZ
4The crosshairs can be hidden by right click on the chosen projection Click on “Hide crosshair”. This will need to be done in each projection you desire the crosshairs to disappear.
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The orthogonal sectioning is synchronized with the movie player, the z-slider as well as with the zoom in functions. Leica Microsystems Inc. 410 Eagleview Blvd, Ste. 107 Exton, PA 19341
Telephone (610)-321-0460 Toll Free 866-830-0735 Fax (610)-321-0425 www.confocal-microscopy.com
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Confocal Application Notes Vol. 4
August 2006
• Auto-Scaling: Auto-Scaling will automatically stretch the image intensity, allowing a better contrast on the screen. It will be applied per channel; hence the auto-scaling is channel dependent. It can be remove by unchecking the Auto-Scaling button, or by using the Reset button in the Histogram function.
Viewer
Auto-Scaling
Auto-Scaling OFF
Auto-Scaling ON
Leica Microsystems Inc. 410 Eagleview Blvd, Ste. 107 Exton, PA 19341
Telephone (610)-321-0460 Toll Free 866-830-0735 Fax (610)-321-0425 www.confocal-microscopy.com
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Confocal Application Notes Vol. 4
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August 2006
Histogram:
This image scaling allows image optimization while in live acquisition. It can be used to enhance dim specimen or where only weak light intensity can be applied (due to phototoxicity or weak fluorescence). Because it is a channel specific scaling, you can balance intensities in different channels. This approach can be also used in fixed cells or tissues. 1- Select the channel. 2- Adjust the histogram slider(s). The vertical slider will move automatically. 3- Adjust the Gamma if needed. 4- You can reset at any moment to get back the original values. This button will reset the Gamma values too. 5- Offline: you can have an automatic rescaling of the min/max values of the LUT in xyz or xyt series (Scale Stack). It will apply the adjusted values to the whole series.
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BEFORE OPTIMIZATION
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PS1: You can use the automatic scaling in conjunction of the histogram tool. In this case the histogram sliders will be blocked. Also, a click on an overlay image won’t show an histogram when the Histogram button is chosen. PS2: Note that the histogram won’t appear when the overlay image is selected (as shown in this image). Leica Microsystems Inc. 410 Eagleview Blvd, Ste. 107 Exton, PA 19341
AFTER OPTIMIZATION
Telephone (610)-321-0460 Toll Free 866-830-0735 Fax (610)-321-0425 www.confocal-microscopy.com
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