ImageJ这套软件可以自动帮你你计算细胞数,也可以定量分析DNA电泳或是Western blot条带。
step 1.首先打开软件后,开启图档ImageJ这套软件可以自动帮你你计算细胞数,也可以定量分析DNA电泳或是Western blot条带。
step 1.首先打开软件后,开启图档step 2.请先做校正,选择Analyze底下的Calibrate选项,再选择校正的模式,使用Uncalibrate OD,再按ok按下ok之后会出现校正的图形Step 3.在要分析的第一条(first lane)加上一个长型框(工具列第一个选项),再按下Analyze/Gels/select first Lane快速键(Ctr+1),此时框架中会出现一个号码1,之后可以移动框架到第二个lane再选择Analyze/Gels/select second Lane快速键(Ctr+2),当然可以一直加下去,最后按Analyze/Gels/plot Lanes快速键(Ctr +3)。
Step 4.分析以后会出现图型表示你刚选择的框内的影像强度,此时可以看到有几个比较高的区段,就是我们想定量的band,使用直线工具(工具列第五个选项)先将图形中高点为有band的区域和没有band的区域分开再,使用魔术棒工具(工具列第八个选项)点选要分析的区域。
Step 5.当我们点选分析时,在result的对话视窗会出现分析的数据,依序点选就会出现每个band的值。
注:当我们选择分析的条带也可以是横向选取,就可以只比较相同大小的DNA 的含量,同样也可以应用在western blot或其它类似实验条带的分析上。
使用ImageJ 分析图像中的颗粒数[] 原创教程,转载请保留此行1,到本站资料下载-实用小工具栏目下载 ImageJ 并安装。
2,打开ImageJ并打开要分析的图片。
请看演示图片。
3,把图像二值话或者设定阈值。
选择Image - Adjust - Threshold...根据提示设定你需要的阈值。
这一步非常重要,关系到结果的正确性。
设定阈值的标准就是把是颗粒的地方都突出出来。
示例中颗粒都被染成了红色。
4,菜单Analyze - Anelyze particles... 自动统计结果。
示例图片中有2个1像素的点都被统计出来了。
结果非常准确。
ImageJ 功能强大,其他功能请自学或者听下回分解!再见~~~ #1使用ImageJ 分析图像中的颗粒数[] 原创教程,转载请保留此行1,到本站资料下载-实用小工具栏目下载 ImageJ 并安装。
2,打开ImageJ并打开要分析的图片。
请看演示图片。
3,把图像二值话或者设定阈值。
选择Image - Adjust - Threshold...根据提示设定你需要的阈值。
这一步非常重要,关系到结果的正确性。
设定阈值的标准就是把是颗粒的地方都突出出来。
示例中颗粒都被染成了红色。
4,菜单Analyze - Anelyze particles... 自动统计结果。
示例图片中有2个1像素的点都被统计出来了。
结果非常准确。
ImageJ 功能强大,其他功能请自学或者听下回分解!再见~~~附件:particle.JPG(9.26 KB)adjust.JPG(26.63 KB)adjust2.JPG(22.57 KB)analyze.JPG(24.67 KB)result.JPG(38.38 KB)2007/4/6 21:53生物医学影像分析软件--ImageJ计算细胞数2009年04月21日 22时21分52秒 来源:未知 浏览: 522次相信许多朋友也许也在找一种影像软体,可以自动帮你计算细胞数,比方说常常用使用4'-6-Diamidino-2-phenylindole (DAPI),或是用其它的抗体在做免疫染色(Immunohistochemistry Staining),要如何把影像的资讯数量化呢?最常用的就是计算染色的细胞数。
那有没有软体可以自动帮你算细胞数呢?大概在90年中就有人在做这种影像分析,在2000年左右也有不少文章介绍各实验室自己开发的软体,也免费的提供下载。
最近由猪排饭同学介绍一套软体ImageJ,是由National Institutes of Health (NIH)所开发免费且跨平台(windows、Mac、Linux都可用)的影像分析软体,试用一下觉得相当好用。
请参考以下之教学:step 1. 下载安装ImageJ软体: /ij/download.html 或ImageJ下载/html/protocol/ruanjianjiaocheng/2009/0421/284.html请依据不同作业系统选择适合的版本step 2. 打开软体,再开启要分析的图片(File->Open)。
step 3.设定分析影像的阈值(Image->Adjust->Threshold,或快捷键Ctr+Shift+T)step 4.调整阈值再按set决定。
step 5.选择analyze particle(Analyze->Analyze particle)并且记得要选择summarize,才会有分析的结果。
step 6.分析结果,可以看到原图像的视窗会出现电脑判读的数据,summary会显示个数及分析影像的区域大小Using ImageJ to Quantify Gel ImagesThis is a quick tutorial abour using ImageJ to process gel images taken with the GelDoc. ImageJ is a free program that was originally written at NIH. ImageJ is used to analyze/process all sorts of images in biology-related research. You can use ImageJ to crop/invert/rotate/enhance your gel images. There is even a way to quantify your bands.Rotating/Cropping Gel ImagesOpen ImageJ using the shortcut on the desktop.You can drag the image you want to open onto the ImageJ window. Once the gel image is open, you can zoom in with "Ctrl+" and zoom out with "Ctrl-". Your gel will look something like this (see below).C:/…/index.html1/10Now you can rotate the image in case it's crooked. Go to Image/Rotate/Arbitrarily... You will get the following dialog window. You can adjust the rotation angle (positive or negative values). To make it easy, make sure that the "Preview" box is checked, that will show you how your image is rotated, and it will also show you vertical/horizontal lines for orientation. Click OK when you're done.C:/…/index.html2/10Now it's time to crop the gel image. Use the "Rectangular Selection" tool. After selecting the region of interest go to Image/Crop to crop the selection.See below for screenshots.Enhancing the Gel ImageThis is a typical step when dealing with gel images. You need to adjust the histogram of the image. Please make sure not to blow-out (saturate) the whites. You want to make sure your image has enough dynamic range. Talk to me if you're confused. Anyways, you can do that withImage/Adjust/Brightness-Contrast... , see the Brightness&Contrast window and the enhancement results below. Click on "Apply" and close the menu.C:/…/index.html3/10Inverting the ImageIn most cases you will want to invert the EtBr gel images for convenient viewing and economic printing. Do this by going to Image/LookupTables/InvertLUT which will give you the result shown below. You can use Brightness&Contrast enhancement once again if you wish, but this one is good enough to move on.C:/…/index.html4/10Quantifying Gel Lanes with ImageJWarning - this is not going to be accurate to the percent. It works and you can put some numbers on your results, but digital camera images are not going to be numerically as accurate as a Typhoon scan. You need to select your lanes first. I will do horizontal selections, but you can also do vertical selections if needed. First of all, click on the "Rectangular Selection" toolDo a tight selection around the lane of interest. Press "Ctrl+1" to designate the first lane. Grab the selection and drag it down to select the background (to use late for background subtraction). Press "Ctrl+2" to designate the second lane, then if you select a third lane, you press "Ctrl+2" for any additional lane. Once you're done, press "Ctrl+3" to plot the lane grayscale density. You can access C:/…/index.html5/10other gel-related functions in the Analyze/Gels menu. If you select horizontal lanes like I did here, you will be asked whether you really want your lanes horizontal. Just say yes.I only had two lanes - the bands of interest and the background lane. So ImageJ plots two separate grayscale profiles. You can see that subtracting the background will really make a difference. If your profiles look weird, please go to Analyze/Gel/GelAnalyzerOptions and make sure that the "Invert Peaks" option is checked.C:/…/index.html6/10Now you will have to manually select your bands. This is done with the line select tool (press shift C:/…/index.html7/10key while drawing a vertical line). This is not as sophisticated as the Typhoon Software, but it offers a bit more control.Now you can identify each band by using the "Wand Tool" (see below). Click with the Wand Tool inside each selection corresponding to a band. Then you can go to Analyze/Gels/LabelPeaks and ImageJ will label each selection defined with the WandTool. There is also a "Results" window that lists the results. Use the "Area" values and make sure to subtract the corresponding background offset value.C:/…/index.html8/10Saving and PrintingAt the end, please save your gel image by going to File/SaveAs, I would recommend saving as TIFF or PNG (lossless). You can email the gel images to yourself. Currently there is no way to print gels directly from the GelDoc computer.Gao yanImageJ Features⏹Runs Everywhere:⏹ImageJ is written in Java, which allows it to run on Linux, Mac OS X and Windows,in both 32-bit and 64-bit modes.⏹Open Source:⏹ImageJ and its Java source code are freely available and in the public domain. Nolicense is required.⏹Plugins:⏹Extend ImageJ by developing plugins using ImageJ's built in text editor and Javacompiler. More than 500 plugins are available.⏹Analysis:⏹Measure area, mean, standard deviation, min and max of selection or entire image.Measure lengths and angles. Use real world measurement units such as millimeters.Calibrate using density standards. Generate histograms and profile plots.Installation ⏹Linux⏹Mac OS X⏹Mac OS 9⏹WindowsWindows/ij/download.html Download ImageJ 1.44 bundled with 32-bit Java 1.6.0_20(28MB), with 64-bit Java 1.6.0_20(24MB; requires 64-bit Windows) or without Java(3MB).OpenReads an image and displays it in a separate window. Files must be in TIFF, GIF, JPEG, DICOM, BMP, PGM or FITS format. Also opens ImageJ and NIH Image lookup tables (with ".lut" extension). Additional file formats are supported via plugins installed in the Import submenu.Subtract background U1幻灯片 10U1 User, 2011/4/17Merge channelScale barScale bars should be present on all publication/presentation images/movies. It’s worth putting them in sooner rather than later. Choose a standard scale bar size for all your images if possible to avoid confusion.⏹If you know the size of a feature (previously applied scale bar for instance) you can use this command to apply a calibration.⏹Using the line selection tool, draw a ling along the length of the feature/scale bar.⏹Run the menu command “Plugins/Spatial calibration/Set scale”[1].⏹Enter the dimensions of the object/scale bar in the “known distance”box and set the units in the Unit of length box.⏹Do not check Global unless you wish all your images to have this calibration! Click OK.⏹。