Please help with Histopathology

[Bernoull]

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Hi,

I'm currently doing research on histology staining and diffuse reflectance spectroscopy. I need help identifying basic structures in stained murine cardiac sections - specificcally mitochondria, nucleus and the margins of the cardiomyocytes (endomysium etc). The sections include normal and infarcted tissue (infarction is secondary to coronary artery ligation) that have been formalin fixed, paraffin embedded and stained via Cain's method (aniline acid fuchsin/methyl blue for primary and counter stains respectively). The sections are 4um thick.

My research has ground to a stop because I can't interpret my findings so I'll greatly appreciate any help you can offer. I've attached some pictures and a copy of the staining protocol.

Thanks for your time and help!

Unfortunately, I wasn't able to attach the pics to this post due to size incompatibility (something about dimensions), I'll try to troubleshoot this. I've also uploaded the pics to google's picasa but I can't figure out how to make them publicly accessible so that i won't have to invite people. I'll work to iron out these IT stuff, in the meantime if someone's willing to help and doesn't mind giving out his/her email (via post or pm), i can add him/her to the picasa album so that the album is accessible to them.

Thanks again, sorry for the complication.

 

[Bernoull]

Hi,

I'm currently doing research on histology staining and diffuse reflectance spectroscopy. I need help identifying basic structures in stained murine cardiac sections - specificcally mitochondria, nucleus and the margins of the cardiomyocytes (endomysium etc). The sections include normal and infarcted tissue (infarction is secondary to coronary artery ligation) that have been formalin fixed, paraffin embedded and stained via Cain's method (aniline acid fuchsin/methyl blue for primary and counter stains respectively). The sections are 4um thick.

My research has ground to a stop because I can't interpret my findings so I'll greatly appreciate any help you can offer. I've attached some pictures and a copy of the staining protocol.

Thanks for your time and help!

Unfortunately, I wasn't able to attach the pics to this post due to size incompatibility (something about dimensions), I'll try to troubleshoot this. I've also uploaded the pics to google's picasa but I can't figure out how to make them publicly accessible so that i won't have to invite people. I'll work to iron out these IT stuff, in the meantime if someone's willing to help and doesn't mind giving out his/her email (via post or pm), i can add him/her to the picasa album so that the album is accessible to them.

Thanks again, sorry for the complication.

I finally figured out how to make the pics publicly accessible without email invitations. Please view pics and help me ID mitochondria, nucleus and margins of cardiac myocytes. Thanks in advance!!!


http://picasaweb.google.com/abuszl1/DropBox#

 

[pathstudent]

I finally figured out how to make the pics publicly accessible without email invitations. Please view pics and help me ID mitochondria, nucleus and margins of cardiac myocytes. Thanks in advance!!!


http://picasaweb.google.com/abuszl1/DropBox#

First. What you going to do for me.

 

[oldfatman]

First. What you going to do for me.

No need to haze the premeds. We were all there once.

Perhaps a good histology book would help you to do it yourself. I thought Wheater's was good when I used it. It had lots of color pictures.

 

[Bernoull]

I'll look around for a Wheater's. I can make out some rudimentary features like erythrocytes, branching cardiomyocytes and perhaps intercalated disks. I know the nucleus is supposed to be centrally located and for the most part uni-nucleated (unlike skeletal muscle). Sometimes I see things i think are nuclei, but without knowing the margins of the cardiomyocyte, I can't tell how many nuclei are present in one cell. I also know the nuclei can be necrotized following the inflammation so focus on only normal tissue until I get a ahng of things..

Having said all this, it'll be great to get a guidance from the experts here on even one slide and I'll use that as reference for.the rest...

To pathstudent: I can offer you some of my student loan debt..lol, that's about all I have to my name. But seriously, I'll be most grateful for ur help.

Thx!

 

[KCShaw]

You'll probably have the best luck if you can find someone locally who can look down the scope with you. Scanning the whole slide, focusing through it, etc., and talking over what you're seeing with someone are all important. Only briefly scanning some of the images, at that magnification you're really only going to be able to structurally identify larger things; nuclei, cell margins, striations, and the like, if not blurred or broken down for some reason. If you're confident of the staining, then one might be able to pick out clusters of mitochondria basically as particles, but the art side of the science starts to come in handy; it's not something people typically go looking for. Is it just me, or do some of the erythrocytes appear blue, and the others red?