- Видео 23
- Просмотров 18 324
Yannick KREMPP
Швейцария
Добавлен 23 окт 2008
This channel lists tutorials on digital imaging for the life scientists.
Original tutorials :
ruclips.net/p/PLK69o_Zliob5YwlaeqoFqCV2ibXWFw57H
Curation from other sources:
ruclips.net/p/PLK69o_Zliob6WMAtKSOgs8mWduB8ciBf8
As a bioimage analyst and instrumentalist, the videos on this channel first and foremost answer questions from users of our imaging platform.
However, if you have a question or an imaging problem, do not hesitate to contact me and I will try to find a solution with you.
[Disclaimer] : My work at the CIF Facility takes precedence to any other activities, so please be patient if you ask a question, I'll do my best to answer it on my free time.
Original tutorials :
ruclips.net/p/PLK69o_Zliob5YwlaeqoFqCV2ibXWFw57H
Curation from other sources:
ruclips.net/p/PLK69o_Zliob6WMAtKSOgs8mWduB8ciBf8
As a bioimage analyst and instrumentalist, the videos on this channel first and foremost answer questions from users of our imaging platform.
However, if you have a question or an imaging problem, do not hesitate to contact me and I will try to find a solution with you.
[Disclaimer] : My work at the CIF Facility takes precedence to any other activities, so please be patient if you ask a question, I'll do my best to answer it on my free time.
CIF Tutorials | QuPath : Building training images for multiplexed analysis + Density maps
CIF Tutorial : This video shows you how to create sparse images to train more robust and unbiased models.
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[Summary]
In this video I show how to create a sparse training image for multiplexed analysis in order to minimize bias. I also show briefly how you can use density maps to create an annotation where most of the triple positive cells are located.
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[Chapters]
00:00 Intro
01:09 Setting up the project, adjusting the display and discovering the samples.
05:59 Creating random annotations to use as building blocks for the sparse image
10:45 Creating the sparse image
14:18 Duplicating the sparse image for each channel
15:25 Cell detection step and saving the command as a script
21:06 Training th...
---
[Summary]
In this video I show how to create a sparse training image for multiplexed analysis in order to minimize bias. I also show briefly how you can use density maps to create an annotation where most of the triple positive cells are located.
---
[Chapters]
00:00 Intro
01:09 Setting up the project, adjusting the display and discovering the samples.
05:59 Creating random annotations to use as building blocks for the sparse image
10:45 Creating the sparse image
14:18 Duplicating the sparse image for each channel
15:25 Cell detection step and saving the command as a script
21:06 Training th...
Просмотров: 1 874
Видео
CIF Tutorials | QuPath : Counting and classifying cells in multiple brain regions
Просмотров 2,6 тыс.2 года назад
CIF Tutorial : This video is an example of how to create a project from A to Z in QuPath. [Summary] In this video I show how to setup QuPath, how to draw regions using various tools, how to detect cells with both cell detetction and StarDist, and train classifiers and create a composite classifier to find positive and double positive cells. [Chapters] 0:00 Introduction 0:44 Creating a project a...
CIF Tutorials | QuPath : Using StarDist with H-DAB images and detections filtering
Просмотров 7752 года назад
CIF Tutorial : How to install StarDist as an extension and use a custom scripts to work with channels created from color deconvolution. We also see how to filter detections that have been measured. [Summary] I quickly show how to install StarDist and download the models, how to navigate the documentation and give a few tips on how to optimize the H-DAB image for analysis. The script used in the...
CIF Tutorials | QuPath : Drawing annotations and arithmetics
Просмотров 7632 года назад
CIF Tutorial : How to use QuPath to draw annotations with a brush and a wand tool, and use some arithmetics to cretae new annotations. PS: I forgot to tell that you need to check [fill annotations] from the view menu to replicate the same appearance, since it's not enabled by default. [Summary] This video shows how to create a full tissue annotation with a tresholder, then create annotations wi...
CIF Tutorials | Imaris : Average distance to structure
Просмотров 962 года назад
CIF Tutorial : How to mesure the average distance between a structure and spots [Summary] In this video I show how to create a surface and edit it manually by cutting and removing unrelevant parts. I the proceed to create spots and keep only those located on the structure of interest and measure the average distance between them and the surface. [Chapters] 00:00 Intro 00:37 Preparing the datase...
CIF Tutorials | CellProfiler : A practical introduction
Просмотров 1622 года назад
CIF Tutorial : How to use start using CellProfiler for basic projects. [Summary] In this short video I show an example on how to setp up a CellProfiler Pipeline from scratch, along a few explanations on how and why to chose certain modules. [Chapters] 00:00 Intro 00:40 Interface and loading images 01:48 Displaying an image 03:26 Using the metada to organize the images 06:13 Naming the images 09...
CIF Tutorials | QuPath : Light Scripting
Просмотров 2242 года назад
CIF Tutorial : How to use QuPath to create regions with a thresholder [Summary] In this short video I'm using the workflow tab to script some actions as a small introduction to how automation works in QuPath. Previous video : ruclips.net/video/Ts5KEh9BH3E/видео.html PS: Sorry for the bad sound... [Chapters] 00:00 Intro 00:30 Cleaning up 01:18 Repeating the processing steps and understanding the...
CIF Tutorials | QuPath : Using thresholders
Просмотров 2792 года назад
CIF Tutorial : How to use QuPath to create regions with a thresholder [Summary] In this video I'm using QuPath with a bright field image to illustrate how to create annotations based on a thresholder. [Features used] - Thresholder [Images] Image from idr.openmicroscopy.org/webclient/img_detail/11908433/?dataset=12438 [Author] Yannick Krempp, bioimage analyst at the CIF platform, Lausanne, Switz...
CIF Tutorials | QuPath : Using multiplexed analysis with superpixels
Просмотров 1,8 тыс.2 года назад
CIF Tutorial : How to use QuPath to create regions with superpixels and classify them like in a multiplex analysis [Summary] In this video I'm using QuPath with a fluorescent slide to simulate a multiplexed analysis on regions created via superpixels. The goal is to label these regions according to criteria more complex than just a simple tresholder and get information for each class and sub-cl...
CIF Tutorials | Imaris : Cell module vs masking + spot tool for vesicle detection
Просмотров 1942 года назад
CIF Tutorial : How to detect vesicles in a subset of cells in a multi-channel dataset [Summary] In this video I show briefly how to use the Cell module to detect vesicles in a cell, and an alternative way using masking techniques and spot detection. PS: Sorry for the very bad audio. I will make sure subtitles are ok. [Chapters] 00:00 Intro 00:20 Viewing the dataset 01:10 Using the cell module 1...
CIF Tutorials | Imaris : Spine detection on a Golgi-cox prepared sample
Просмотров 2632 года назад
CIF Tutorial : How to convert and prepare a Golgi-Cox dataset for spine detection. [Summary] Golgi-Cox preparation allows for very nice filament and spine labeling, but you have to do some light prepocessing in order to use the data in Imaris. In a second part, I show how to trace dendrites and detect spines using a semi-automatic way by working from selected somas. [Features used] - Imaris XTe...
CIF Tutorial | Imaris : Manual filament tracing and soma import from Surface
Просмотров 7042 года назад
CIF Tutorial | Imaris : How to create an Imaris fialement object manually with a surface object as soma. [Summary] In this video I show how to quickly create sevral surfaces from a complex dataset using a semi-automatic way. For difficult cases there is also the option to go fully manual, which is also shown. Once created, these surfaces can be imported into a new Imaris Cell object that repres...
CIF Tutorial | Fiji | Segmenting Muscular Fibers
Просмотров 4,7 тыс.3 года назад
CIF Tutorial | Fiji | Segmenting Muscular Fibers [Summary] How to use Fiji along with MorpholibJ and StarDist to segment muscular fibers from a color slide scanner image. NB : No method is perfect and this one is no exception. It can obviously benefit from further refinements like better filtering, and maybe manual label edition to correct obvious mistakes. There are also other methods involvin...
CIF Tutorial | QuPath | 003 - Using Stardist with QuPath built from source on Windows
Просмотров 1 тыс.3 года назад
CIF Tutorial | QuPath | 003 - Using Stardist with QuPath built from source on Windows NB: Optional step for advanced users [Summary] Once you have built your custom QuPath version, you can use Stardist to detetc cells. This video shows you how to apply these kind of scripts. [Notions viewed] - Cell detection and classification - StarDist - Scripting [Images] NC [Author] Yannick Krempp, bioimage...
CIF Tutorial | QuPath | 002 - Building QuPath from the source code
Просмотров 1593 года назад
CIF Tutorial | QuPath | 002 - Building QuPath from the source code. NB: Optional step for advanced users [Summary] To be able to run scripts using tensorflow like StarDist, you need to compile your own version of QuPath. In this tutorial I will show you how to do this on a Windows PC. [Notions viewed] - Getting source code from Github - Using Gradlew to compile QuPath [Images] NC [Author] Yanni...
CIF Tutorial | QuPath | 001 - Installation
Просмотров 2543 года назад
CIF Tutorial | QuPath | 001 - Installation
CIF Tutorials | Manual and semi-automatic surface creation
Просмотров 573 года назад
CIF Tutorials | Manual and semi-automatic surface creation
CIF Tutorials | Imaris : Count and classify spots in a masked region
Просмотров 6653 года назад
CIF Tutorials | Imaris : Count and classify spots in a masked region
CIF Tutorials | Imaris : Filtering and resampling for better 3D reconstruction
Просмотров 8203 года назад
CIF Tutorials | Imaris : Filtering and resampling for better 3D reconstruction
CIF Tutorials | Imaris : Detect, filter and classify spots
Просмотров 3703 года назад
CIF Tutorials | Imaris : Detect, filter and classify spots
CIF Tutorials | Imaris : Prepare your data for the analysis
Просмотров 1483 года назад
CIF Tutorials | Imaris : Prepare your data for the analysis
CIF Tutorials | Imaris & the Cell module
Просмотров 1753 года назад
CIF Tutorials | Imaris & the Cell module
HRM 001 - Deconvolution : Using the Huygens Remote Manager (HRM) at the CIF platform
Просмотров 1414 года назад
HRM 001 - Deconvolution : Using the Huygens Remote Manager (HRM) at the CIF platform
Hi Thank you so much for such an amazing video. I am trying to use this video for my cells analaysis. However I am not able to run classifier to detect particular channel. Could you please tell me what should I do
I assume you are trying to use StarDist? If yes look at the code part halfway through this video (e.g at 21:30). At some point you have to specify on which channel you want to work with. In this example it's 'DAPI'. It may be different for you, you can look for the channel names in the Brightness/Contrast tool (the half black circle in the toolbar next to the drawing tools). If you are asking about running a classifier you have trained, make sure you apply it on objects detected in the same way as the objects used for the training. For instance the StarDist script doesn't always include the "measureIntensity()" command, meaning a classifier trained on detections which have an Intensity feature will not work on detections that have not this feature. One common issue is that the Analyze>Cell detection command adds the intensity features by default, but not the StarDist code snippet. Another common mistake is trying to apply a classifier trained on images with certain channel names on images which have other channel names (for instance, DAPI became Alexa405nm or something similar). There are many potential issues like that, the key being to try your best to stay consistent.
Thanks for the great and valuable tutorial! Really appreciate it!
Thanks! very useful! There is an option to change the radius of the detecting cells with Stardist?
All the available parameters are listed here: qupath.readthedocs.io/en/stable/docs/deep/stardist.html You can change the cell expansion radius, but not the nuclei. I suppose you could filter out later nuclei out of your target range. Something like that: forum.image.sc/t/select-delete-objects-that-are-above-a-specific-area/38229
Nice!
Is it possible to segment nuclei with StarDist, after using a Colour Deconvolution for H/E or from original H/E image? I have triend with both images as input but it looks like StarDist segment preferably larger objects (i.e. muscle fibers) rather than nuclei. The result is that lots of nuclei are included in the muscle fiber object. This happens to me also if I use Hematoxylin image, obtained after using Colour Deconvolution. Thank you for your tutorial!
Stardist is quite sensitive to the cell (or round object) size. A good trick is to scale the image according to what stardist expects. Example of the issue here with a blob image scaled up 8x : ibb.co/hLh9Hch . Another way to improve woud be perhaps to try to do a better deconvolution. If not possible, try to resort to pixel classifiers plugins like weka or labkit to create a mask of the regions you want to exclude, then use stardist on the masked copy of the image
@@UniversalBuilder Thank you for the suggestions! 😎
Hey Yannick, many thanks for your help and efforts!! Could you please kindly tell me what kind of staining technique you're using in the sample image?
This wasn't my sample so I'm not 100% sure but this was probably a simple hematoxylin and eosin labeling.
Can you please tell what magnification image you have selected here?
If you are referring to the acquisition with the slide scanner it was probably the 10x plan apo
Nice video, but can you please be louder since there are no captions it is hard to understand
Sorry for that. Since then I invested in a decent mic and the sound should be much better in the next videos. I'm a bit short of time these days so I had no updates for a long time but I'm planning to get back on track soon. Thanks for watching !
very nice, thank you!
Great video! I would like to know How do you determine the parameters of Stardist?
Thanks ! You can find some answers here : imagej.net/plugins/stardist And there's a FAQ (that also applies to training models and the python version) here stardist.net/docs/faq.html In general, start with the suggested parameters for the model. If it doesn't work well, try first to check the size of your images to see if they need to be resized to get a proper segmentation. The rest depends on your images, so you might have to check one parameter at a time and see how changing it improves or not the results.
Thank you very much for sharing such an informative tutorial. One of the best thing for me was the easy step-wise explanation for every sub-step. Keep on this good work
Thank you for this super useful tutorial!
Very useful tutorial.
Hi Thanks to this video tutorial. Can we do this Segmentation in 3D cultured cells and if yes please make one video on 3D cultured cells Segmentation.
Hi, it's possible but I wouldn't suggest using Fiji for that. I would use something like Imaris, or maybe try other tools like APEER, ... In order to enable 3D segmentation with StarDist and Fiji you have first to install and configure a proper Python environment and the StarDist full package. Then you can use a wrapper to connect from Fiji to your Python installation. There are instructions here : github.com/BIOP/ijl-utilities-wrappers The rest of the workflow should be quite similar. As of making a video for that, I'll see if I have time but no promises there, as I'm really too busy in the moment :(
@@UniversalBuilder Thanks for your suggestion I will try to do
Technobear suggested an alternative method using Points instead of using the brush to create annotations for the training. This workflow works well, but only if you detect the cells BEFORE splitting the sparse image into separate channels. You can then directly enable the Point tool at this step. This method is faster than what I shown, so give it a try and tell me if it's useful to you. I can give more explanations at the Image.sc forum which is better suited to show code and screenshots : forum.image.sc/t/thank-you-for-the-youtube-tutorials/68833 Thanks Technobear
Hello, Thankyou for the video. Wanting to confirm if the label part at the end is the number of muscle fibres?
Slightly late to answer (sorry didn't see this one), Yes it's the number of particles analyzed.
Thanks for this video! Would it be possible to make a video to install the QuPath-Tensorflow version on windows computer please ?
Thank you ! I will have a look at this question, but I'm currently waiting the release of the 0.4 version to check if there's something new on that side. If not, I will record a video to show how to do that from scratch
@@UniversalBuilder Thanks a lot!
Hello Marion, just a quick update: QuPath 0.4 is finally out! Regarding Tensorflow, it's as I suspected: everything was rebuilt and the past extension is now obsolete. Building QuPath from scratch isn't needed anymore apparently, or at least it seems much easier. Please look at this thread from the developper : forum.image.sc/t/qupath-v0-4-0-now-available/74887. Good stuff :)
@@UniversalBuilder hello Yannick, thanks for your commentary! I have also seen this new version and it is much easier to use Stardist with Tensorflow and that is a good thing!
Thank you very much for the tutorial. Do you have any advice for automated quantification for IHC-stained muscle fibers? I have to count the positive fibers(not cells) and have been doing it manually. Thank you in advance!
I think you could probably do a variation of the tutorial above and add another step to discriminate between the fibers, but it's difficult to say without knowing what you consider as "positive". Is it a change of size or shape ? Of staining color ? Of texture ? Depending on that you could use tools to get information from the ROIs created using this tutorial to create a new heat map based on the measurements, and threshold it to keep only the fibers you estimate as positive. The Biovoxxel and MorpholibJ plugins have tools for that if I recall correctly.
@@UniversalBuilder , thank you very much for your response!
so good.