Compara've Medicine Animal Histology Services. Glossary of Terms

Transcription

1 Compara've Medicine Animal Histology Services Glossary of Terms

2 About this guide This guide is intended to provide clarifica'on on standard histologic terminology. Included are examples of different sample types, processes, and final results Types of samples submi/ed by the inves5gator Processes available Final result depending on the process chosen Remember to always include a request/order form with every request. Failure to do so will cause delays in processing of your request(s).

3 Star'ng samples This sec'on provides informa'on on what your star5ng sample is. The star5ng sample is what you will submit to the histology lab for processing of your order/ request.

4 Fixed and Unprocessed Tissues These are 'ssues that have been fixed, trimmed, and placed in 'ssue casseges. These 'ssues are submiged to the histology lab in 70% ethanol. Please make sure that all containers are leak- proof and 5ghtly sealed. Star9ng Samples

5 Fixed and Unprocessed Tissues ** Ensure Assue cassebes are clearly labeled with pencil or solvent- proof ink, and are placed in a 9ght- sealing, leak- proof container clearly idenafied with your name, date, PI, & sample IDs. Always include a request form! Images: h>ps://cvmbsresearch.wordpress.com Star9ng Samples

6 Paraffin Tissue Blocks These are 'ssues that have already been processed and embedded in paraffin (wax) to form a 'ssue block. These blocks should be submiged in a container or zip lock plas'c bag. Please make sure that all containers are closed to avoid losing blocks in your submission. Star9ng Samples

7 Paraffin Tissue Blocks ** Ensure Assue blocks are clearly labeled, and are placed in a closed or sealed container clearly idenafied with your name, date, PI, sample IDs. Always include a request form! Image: h>p:// Star9ng Samples

8 Unstained Slides These are slides that have been made from paraffin (wax) 'ssue blocks sec'oned with a microtome, and are intended for later use such as rou'ne, special, or immunostaining. These slides should be submiged in a plas'c slide carrier that keeps slides separated. Please do NOT submit slides loose inside a container. This can result in the paraffin sec5ons being scratched off the glass slide if the slides rub against each other. Star9ng Samples

9 Unstained Slides ** Ensure glass slides are clearly labeled, and are placed in a container that keeps slides separated and is clearly idenafied with your name, date, PI, sample IDs. Always include a request form! Images: h>p://library.med.utah.edu/ Star9ng Samples

10 Frozen OCT Tissue Block These are 'ssue blocks of frozen 'ssue than have been embedded in OCT (Op'mal CuUng Temperature) medium. These blocks must remain frozen and not be exposed to thawing. Please contact the laboratory personnel for coordina5ng the transfer and handling of frozen OCT blocks. Star9ng Samples

11 Frozen OCT Tissue Block ** Ensure plasac OCT molds are clearly labeled. ** Please contact the lab personnel for the transfer and handling of OCT frozen Assue blocks. Images: h>p:// Star9ng Samples

12 Slides from Frozen OCT Blocks These are unstained slides obtained from frozen OCT 'ssue blocks that can be used for a variety of processes including rou'ne, special, and immunostaining. Please contact the laboratory personnel for coordina5ng the transfer and handling of slides from frozen OCT blocks. Star9ng Samples

13 Slides from Frozen OCT Blocks ** Please contact the lab personnel for the transfer and handling of slides from OCT frozen Assue blocks. Image: h>p:// Star9ng Samples

14 Services Offered and Request Types This sec'on provides informa'on on what types of histology services are available for you to request. Please be sure to always include a completed request form with your submission. Failure to submit your REQUEST FORM will result in delays.

15 Tissue Processing ONLY Processing involves a series of 'ssue dehydra'on, clearing, and paraffin infiltra'on steps. Processing prepares 'ssues for paraffin embedding. Final result: Paraffin- infiltrated 'ssues in casseges (but that have NOT been embedded into blocks). Services, Requests, and Final Results

16 Tissue Processing ONLY ** Tissue cassebes are processed in the Assue processor. APer processing, Assues are infiltrated with paraffin and are ready for embedding. Images: h>p:// path.com/ h>p:// Services, Requests, and Final Results

17 Tissue Processing and Embedding ( Blocking ) This service includes embedding of previously- processed 'ssues into paraffin (wax) blocks. Paraffin blocks can then be sec'oned with a microtome for making slides. Final result: Paraffin 'ssue blocks. Services, Requests, and Final Results

18 Tissue Processing and Embedding ( Blocking ) ** Processed Assues are embedded in paraffin and made into paraffin Assue blocks that can be secaoned with a microtome. Images: h>p://9ssuesampling.weebly.com/ h>ps://liferangroup.org Services, Requests, and Final Results

19 Recuts and Step Sec'ons Recuts are sec'ons made from paraffin blocks that have been previously sec'oned. Step secaons are recuts that are obtained a certain distance from the face of the block and from each other. For example, you may order 3x 50um step sec'ons. This means that the second recut will be 50um deeper than the first one, and the third recut will be 50um deeper than the second one. These are also referred to as levels or measured levels Final result: Slides containing a paraffin 'ssue slice. Recuts or steps can be requested as stained or unstained slides (see Unstained Slides sec5on next). Services, Requests, and Final Results

20 Recuts and Step Sec'ons ** These are examples of serial or consecuave recuts. Paraffin blocks are secaoned with a microtome and then collected onto glass slides. Images: h>p:// Services, Requests, and Final Results

21 Unstained Slides Unstained slides can be used for further rou'ne, special, or immunostaining. Unstained slides can be requested as serial slides or step sec'ons. For example, you can request 5 unstained serial sec'ons. This means that 5 consecu've sec'ons will be obtained and placed on glass slides Final result: Slides containing a paraffin 'ssue slice. Services, Requests, and Final Results

22 Unstained Slides Images: h>p://library.med.utah.edu/ Services, Requests, and Final Results

23 H&E Staining Hematoxylin and eosin (H&E) is the principal stain used in histology, and regarded as the gold standard for brigh`ield microscopy. H&E- stained slides provide a context in which to interpret specialized techniques such as special stains and immunostains. Final result: H&E- stained slides. Services, Requests, and Final Results

24 H&E Staining Images: h>p://emedicine.medscape.com/ h>p:// Services, Requests, and Final Results

25 Histochemical (Special) Staining Histochemical (special) stains aid in the visualiza'on and/or iden'fica'on of structures and/or substances in 'ssue sec'ons. Examples: lipid, intercalated discs in cardiac cells, metals and other cell accumula'ons, collagen, infec'ous elements, basement membranes, etc. Final result: *Various* Please consult with the laboratory personnel for iden9fica9on of the most appropriate special stain for your project and research needs. Services, Requests, and Final Results

26 Histochemical (Special) Staining ** Masson s Trichrome for collagen. ** GrocoB s methenamine silver (GMS) for fungi. ** Periodic acid Schiff (PAS) for mucins. ** Perls Prussian Blue for iron. Images: JG Vilches- Moure ** ReAculin stain for reaculin fibers. ** Warthin- Starry silver for spiral bacteria. Services, Requests, and Final Results

27 Immunostaining Immunohistochemistry (immunoperoxidase) assays are based on the principle of an'body binding used to iden'fy specific an'gens and an'genic determinants (epitopes) in 'ssue sec'ons. Examples: different cell types (e.g. B- cell v. T- cell, endothelial cells) cellular components (e.g. mitochondria), specific matrical components (e.g. collagen IV), etc. Final result: *Various* Please consult with the laboratory personnel for iden9fica9on of the most appropriate immunostain for your project and research needs. Services, Requests, and Final Results

28 Immunostaining CK8 Ki67 Images: JG Vilches- Moure Services, Requests, and Final Results

29 Compara've Medicine Animal Histology Services For addi'onal ques'ons or inquiries, please contact: Dr. José G. Vilches- Moure Veterinary Pathologist Director, AHS Dr. Kerriann M. Casey Veterinary Pathologist Dr. Donna M. Bouley Veterinary Pathologist