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发表于 2011-11-1 21:39:19
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chujindong 发表于 2011-11-1 23:28
那就是弄不成了，在DIVA上做的如果需要这种图的话还得拿到flowjo上去做。
并非全无办法。见这篇文章：http://www.ctt-journal.com/index.php?id=255
Part 2: How to generate histogram overlays using GIMP (based upon Part 1)
If a histogram overlay needs to be created, the respective plots have to be pasted into and processed within a graphic-editing program able to create overlays, such as GIMP. First, copy histogram 1 to the clipboard using Acrobat Reader and paste it as a new image into GIMP. To do so, open GIMP and click File --> Create --> From Clipboard in the menu bar.
Now switch back to Acrobat Reader and copy histogram 2 to the clipboard (which has ideally been set to another color in FACSDiva beforehand). Make sure both histograms are of the same size (see step one); this is essential. Histogram 2 has to be pasted as a new layer into the same image file of GIMP. To do so, click Edit --> Paste as --> New Layer.
The image file now contains the two histograms lying on top of each other in two Layers. In the layers dialog of GIMP (usually at the right side, otherwise press Ctrl+L) the two layers are shown as well as the Layer Mode (set to Normal as default). The mode determines how the layers interact with each other and may be used to combine the two histograms into one overlay graph. To this aim, set the layer Mode to Multiply. Now both layers are visible at the same time, but not yet aligned. To align both histograms, set the Zoom function to 100% (View --> Zoom --> 100%). Select the Move Tool (Tools --> Transform Tools --> Move), click on one of the histograms and use the cursor keys of your keyboard to move it until both histograms are fully aligned.
Set the Zoom back to about 33% (View --> Zoom --> 33%) to see the entire histogram overlay. If the two histograms had different titles, the overlay may result in an unreadable text mixture. This can be initially avoided in FACSDiva (by applying identical or no titles), or corrected in GIMP, e.g., by using the Eraser Tool (Tools --> Paint Tools --> Eraser).
To save the image, click on File --> Save as, enter a filename such as Overlay1.tif to save the file as a TIFF image. Now select a folder and click OK. In the next dialog choose merge visible layers, and then check LZW to use a lossless compression for the TIFF-file. This saves space without reducing image quality. In the supplemental material hints are given how the design of histogram overlays could be altered.
In Figure 2, an example is provided in which a fusion protein consisting of EGFP and ZeoR has been expressed in 293T cells using LeGO vectors [4]. The histogram overlay shows transduced cells before and after selection with zeocin.
Figure 2. Straightforward composition of histogram overlays. 293T cells were transduced with a LeGO vector [4], encoding a fusion protein consisting of EGFP and ZeoR (Weber et al., submitted). Gene transfer reached 10%, i.e., 90% of the cells remained EGFP-negative (dark grey histogram). After selection with Zeocin, almost all remaining cells were EGFP-positive (99.8%; green histogram).
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发表于 2011-11-1 21:39:18
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