For image analysis, as these images are acquired with the same setting on the same day, should all images use the same brightness/contrast for proper comparison? I asked this because it turned out that the intensity of the same fluorochrome-conjugated antibody can be very different among different samples, which makes me hard to choose a single brightness/contrast that can fit all samples (some samples will be overexposed or underexposed, which to me do not make much sense to analyse such data). Would it be possible to use Auto-brightness/contrast for all samples, and do comparison among them?

EdmundOptics Catalog

Well if you want to segment all you samples automatically, your goal is to find a processing pipeline that give you good results every time (or almost) with the same settings, this is the fastest and more reproducible way.

EdmundOptics logo

In both cases DOF is measured in front of the focus point and behind the focus point. DOF is determined by three factors – aperture size, distance from the lens ...

Hello everyone, I am new to imaging, and would like to seek consults for proper image analysis. I acquired my images of different samples with Axio Scan using the same setting on the same day.

How about % positive area? Do we still need to create a mask? As we are only interested in obtaining positive signal from the background, does it mean that we can just process images with different settings to obtain the best segmentation in different samples, and measure the % positive area? Basically this will be like measure the positive area in the mask image?

Hi @xhs1314 , Thanks for the sample Yes contrast settings can influence the thresholding in ImageJ IF the ‘‘don’t reset range’’ box is checked and your image is 16-bit or more. So I invite you to uncheck it. No, contrast will not change your pixel values, as long as you don’t click on the ‘‘Apply’’ button in the B&C window!

Thank you for answering. Here I uploaded an image stack of two samples. As you can see, although they were acquired under the same setting, the intensities are very different between them. I am interested in quantifying the percentage of positive area and also fluorescence intensity. Ctr2&4 stack.tif (9.0 MB)

EdmundOptics address

Could you suggest a general pipeline for this kind of image analysis? Other than filter, brightness/contast, and threshold, any more common procedure that people usually do to improve segmentation?

This is an image of a store front on a bright blue sky day. The. Your one stop shop for kitchen gadgets, home decor, and more located in scenic downtown Edmonds ...

20231023 — F-number is defined as the lens focal length (f) divided by the lens aperture diameter (d). It defines the size of the cone of light that is focused on the ...

EdmundOptics careers

Edmund Optics Polished Aspheric Lens image 1. Edmund Optics Polished Aspheric Lens image 2. Updated: 7 hours ago. Polished Aspheric Lens. 25mm Dia 0.63 NA ...

After setting brightness, can I use auto-threshold for each of the sample? It makes more sense to use auto-threshold, as each sample has a different background. Or only one threshold can be used and applied to all samples? One manual threshold, again, will lead to non-specific signals in different samples.

With that said, in order to get a good threshold you are completely free to filter and process a duplicate of your image as much as you want! Once you have a good mask, you can use it to measure fluorescence intensity on your original file. Here with your sample, if you apply on both images a median filter with a radius of 10 and try a default auto threshold (with the box unchecked) you get a quite good result : image1588×1116 94.6 KB

Jul 16, 2021 — Learn what "rack focus" means along with other filmmaking terms and phrases in Filmmakers Academy's glossary...

EdmundOptics wiki

So how should I do to overcome this problem, if my images of different samples show distinct intensities? Auto-brightness? Auto-thresholding? Is this ok?

EdmundOptics revenue

Thanks for the great explanation! So if I understood it correctly, I can do anything I want with the duplicates of images (filter, brightness/contrast,threshold etc), and create a mask and use it to measure fluorescence intensity of original file. So, the settings can be different for image duplicates of different samples, as long as it can segment the positive region out, and reapply to original image, am I correct?

So, the settings can be different for image duplicates of different samples, as long as it can segment the positive region out, and reapply to original image, am I correct?

EdmundOptics headquarters

EdmundOptics locations

If the filter and auto-threshold give you good enough results to answer your biological question, I suggest you keep it simple! If you really need perfect segmentation, you can check other filters or machine learning (like the Labkit plugin) and if everything fails you can try deep learning for the rescue For all of that I invite you to explore this forum, the Imagej.net and this awesome tutorial : Introduction to Bioimage Analysis — Introduction to Bioimage Analysis

Could you suggest a general pipeline for this kind of image analysis? Other than filter, brightness/contast, and threshold, any more common procedure that people usually do to improve segmentation?

Ist Glauben Wiederholbar? (Kierkegaard Studies. Monograph Series, 6) (German Edition) [Beyrich, Tilman] on Amazon.com. *FREE* shipping on qualifying offers.

Before purchasing or using a microscope, it is important to know the functions of each part. Eyepiece Lens: the lens at the top that you look through. They are ...

SOLYX® Light Diffusing Window Films feature a collection of light diffusing films designed to allow light through while cutting glare and protecting ...

Oct 31, 2023 — Each of these numbers corresponds to a unique aperture setting. Lower numbers signify larger apertures, allowing more light into the camera and ...

the brightness and contrast is just about visualizing your image, only the display will change not your pixel values. Could you provide a few sample images in original format so we can help you more efficiently? And please tell us more about what kind of analysis you want to make after segmentation: is it fluorescence quantification? counting? morphology? etc

Although changing the brightness and contrast would not lead to changes in pixel values, the selected brightness and contrast would affect thresholding, which will be necessary for area and fluorescence intensity quantification. Fluorescence intensity quantification will be affected too after changing brightness and contrast.

Wicked ABEC9 Freespin are the best among all ABEC rated models released by the brand. These are great quality, versatile bearings which can be used for ...