You could DWI/MRI or perfusion-weighted MRI as a good measure of the infarct size as a result of ischemic injury...

The previous comment is assuming its an animal model...

depends on what scale you are working at and the anticipated size of the infarct. You could simply take the brain and using a simple guide take serial sections with a razor blade. Take photomicrographs thru a dissecting scope or macro camera lens and proceesed the image with Image J. You could also do some staining, a standard nissl stain will catch the boundries of the infarct. Otherwise you can use any number of standard microscopic approaches, collect a known number of sections, measure with a micrometer and calculate volume, or photograph and use image J for processing. Most silver stains will help you highlight the amount of degeneration you have going on. Now if you want sophisticated staining for any number of genetic factors of proteins....well then it gets complicated.

If the I/R injury is in a rodent model, a commonly used method of measuring infarct volume is through staining the isolated brain with 2% 2,3,5-triphenyltetrazolium chloride (TTC), which labels non-injured tissue, leaving the infarct area white. Cut the brain into ~2 mm coronal segments and take pictures of each sample, and as was mentioned in the previous post, use ImageJ or other similar software to outline and measure the area of infarct for each section. Once you have the infarct area for all sections, you can calculate total volume of the infarct within the brain. I have included a link to a paper by Ren et al., (2003) in PNAS studying the effects of lithium on I/R injury which may better describe the protocol for this staining method.

Ren et al. (2003) Postinsult treatment with lithium reduces brain damage and facilitates neurological recovery in a rat ischemia/reperfusion model. PNAS 100(10) 6210-6215.

http://www.pnas.org/content/100/10/6210.full

Thanks to all. The information given is useful and I have tried the suggestions, but still I find it difficult to make use of ImageJ software. If anyone can provide detailed plan/flowchart as a guide to make use of ImageJ. I have searched web for this and will continue. Your reply is precious to my work.

Hi Vishal. This is actually quite easy, though ImageJ looks pretty intimidating at first. I'll explain this in the experimental case where you are imaging coronal sections and determining the area of the infarct as a percentage of the total brain area or volume. This is the easiest way to present these data.

1) Open ImageJ and click on File -> Open, and open the image of interest. For this it is preferable you have an image where the entire brain fits within the boundaries of the image. If not you may create a montage of single picture of higher power to form the entire brain image that will fit within the image window.

2) From the tabs available above the buttons on the toolbar, select Analyze -> Set Measurements. Make sure Area and any other parameter you are interested in is checked on this screen. The units of area measurement may look confusing as it may be in unit of pixels, but it will not matter for taking the size of the infarct and dividing by the size of the total brain, as this will give you a percentage.

3) Once you have set your measurement parameters, click on the Polygon Selections button on the tool bar. This should be third from the left in recent versions.

4) With your image open, trace the entire brain section by clicking and outlining the perimeter of the brain. Once you have outlined it, click near the starting point and the contour selection of the brain will be complete.

5) Select Analyze --> Measure from the tabs above the toolbar. A Results box should pop-up displaying the Area measurement and any other parameters you selected earlier.

6) Follow this same tracing method for the infarct area of the brain tissue, and again select Analyze -> Measure, and the results block will show the area of the infarct selection, which should be a much smaller number than the whole brain measurement. For a single tissue section just divide the infarct area by the brain area and you will get a Infarct area as a percent of the total brain tissue. You can also do this with just the ibsilateral or contralateral hemispheres if this is how you plan to represent the infarct data.

7) You may do this for all your tissue sections of interest, but my advice is to measure all the total brain areas for all brain sections, as well as for the infarct. Determine the total volume of the infarct and brain, and then present infarct volume as a percentage of total brain volume. This is a very appropriate and simple way of effectively displaying and comparing the sizes of infarcts in an experimental design. Hope this helps!

Thre are also some very "old school" methods that include using a projecting microscope or camera lucida and tracing onto grid paper and then counting squares, or even cutting out the area of interest and weighing them. They do work, if done properly, but I am not sure how journal reviewers would react to that approach.

As Wayne mentioned, these methods work as well. Also, Wayne, you are correct, I am not sure how reviewers would view such techniques today. My recent publication used the methods described in my previous post to assess lesion volume in spinal cord tissue after injury, and these methods were well accepted and were not questioned by reviewers. Camera lucida drawings are very cool to see, though Neurolucida by MicroBrightfield, a powerful hardware/software set up allows you to do it digitally and reconstruct tracing contours in 3D, a method I have also utilized in publication. This is now the upgraded standard in neuroscience research from what I can tell.

Thank you Chandler and Wayne.

My lab operates on the proverbial shoestring budget and I am often diving into some of the more low tech (read cheaper) methods. I have found that as long as there is a reference the journals have been receptive. In fact, I wish there were a web site out there with a collection of some of these older methodologies, some of them are very useful.

Very true, Wayne. I am fortunate to have more resources at hand, but science doesn't require the newest most expensive technology to get published. I still fall back on Nissl and Luxol Fast Blue quite often for some of the important assessments in histology. No one will question these methods unless you are trying to do something outrageous with them. I have gone through many of these older techniques and many are still used in our lab. We have a text coming out due in April that covers most all general and advanced histological, physiological, and behavioral assessments using common animal models in acute neural trauma and disease. This is a follow-up to a previous volume extensively describing the animal models themselves. if at all interested, here is a description and Table of Contents of sorts of the newer volume.

Animal Models of Acute Neurological Injuries II. Chen, J.; Xu, X.-M.; Xu, Z.C.; Zhang, J.H. (Eds.)

http://www.springer.com/biomed/neuroscience/book/978-1-61779-575-6

The first volume, is surprisingly enough "Animal Models of Acute Neurological Injuries".

Oh, and go Lopers! I earned my M.S. degree from UNK. Dr. Hoback was my advisor, when I was doing ecological field research. Small world.

Also, concerning the ImageJ software, the NIH provides it as a free download, and I use it the most for image analysis. It is just a bit difficult to get used to.

Yes, I know Wyatt, how long ago were you at UNK? I am getting my 20 year pin in April. I have used Image J for some time now and there is a bit of a learning curve but it's not too bad.

I will look up your fist volume and have the library order the new one, and maybe the first if It is still in print.

I did my M.S. from '06 to '08. He and a co-author and I are working on a manuscript from research during that time. Glad to still be in touch with him.

Concerning the texts, the first volume is still in print, and is a wonderful reference for anyone, especially rookies, in the field. This was published before I joined the lab, but I still use it for solid reference material. The upcoming volume, I had the great opportunity to contribute a small part to describing morphological assessments following SCI, but is filled with chapters by top researchers in stroke, brain and spinal cord injury, and more.

The answers above covered excellent methods in measuring infarcted tissues. I just want to highlight the need to correct for edema, which is inherent after stroke. The JCBFM paper published more than two decades ago by Dr. Swanson and Dr. Sharp is a very resource for measuring cerebral infarcts and correcting for edema (J Cereb Blood Flow Metab. 1990 Mar;10(2):290-3).

Thanks to all........

U can get various softwares to measure infarction in photose of brain ex. image J software can be used to measure infarcted area in brain sections stained with TTC

I think it is better to measure infarct volume rather than % infarct area, as it is more accurate than % infarct area. For measuring infarct volume use TTC or MRI after MCAO and measure infarct in image J software.

% Infract Area gives you a better idea of Brain Injury due to induction of ischemia in comparison to whole brain

Use Image J software as Dr. Walker Suggested. It's the perfect way to rule out the infract.

Measuring infarct size by the tetrazolium method (southalabama.edu)

A Brief Review of Edema-Adjusted Infarct Volume Measurement Techniques for Rodent Focal Cerebral Ischemia Models with Practical Recommendations - PMC (nih.gov)

Measurement of Infarct Size Using MRI Predicts Prognosis in Middle Cerebral Artery Infarction | Stroke (ahajournals.org)

Cerebral infarct volume measurements to improve patient selection for endovascular treatment - PMC (nih.gov)

Two simple and rapid methods based on maximum diameter accurately estimate large lesion volumes in acute stroke - Kufner - 2020 - Brain and Behavior - Wiley Online Library

ImageJ sofware is suitable for this and Chandler L. Walker sir given detail procedure.

Post Infarction Distal Subtraction Angiography and Serial Diffusion-weighted MRI can be used to identify salvageable vs non-salvageable areas and decide the candidates between thrombolysis and Decompressive Craniectomy for Better Treatment.

Hello Sir, with the help of software on a remnographic imaging(MRI or ST scan) you can easily do this. I used software before surgery the subdural hematoma to measure the volume