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Steps to Better Grossing

Geoffrey Rolls , BAppSc, FAIMS

From patient to pathologist, preparing tissue specimens for histological examination requires care, skill and sound procedures. This guide provides practical advice on best practice techniques and simple ways to avoid common errors.

Tips for better grossing are highlighted in this guide. We hope each step provides a valuable reminder of good histology practice and also helps with troubleshooting when unacceptable results do occur.

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Check Fixation Status

Specimens are dealt with promptly (especially large specimens that may otherwise be inadequately fixed).

No consideration given to optimizing the fixation of problem specimens.

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Step_12_Specimen_pots_various_sizes
Specimens in the large containers should be checked as soon as possible to ensure they will be adequately fixed.

Prepare Thin Slices

Care is always taken to prepare uniform, thin slices from large specimens (3–4 mm maximum thickness). This is particularly important with dense tissues.

Slices are sometimes 6 mm (or more) thick and are often uneven.

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Step_13_Well_trimmed_specimen
Shown here are uniform, thin (2–3mm) slices of a tumor ready for processing. They should process effectively and should section without difficulty.

Avoid Specimen Trauma

Care is taken to avoid traumatizing delicate specimens, particularly those that are incompletely fixed (handle carefully, do not crush, always use sharp blades).

Specimens are handled roughly without any consideration as to their state of fixation. Sometimes blunt blades are used in dissection.

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Avoid Cross-contamination
Section of H&E stained lung showing obvious local trauma due to very forceful grasping with forceps. Fresh or partly fixed tissue is most susceptible to damage, but even well-fixed tissue can be damaged by rough handling.

Avoid Cross-contamination

Each specimen is handled on a clean surface avoiding the possibility of specimen-to-specimen contamination.

Sometimes the surface of the cutting board is not properly cleaned between specimens. This is of concern when the same specimen types are cut up one after the other. You do not want to have carry over from a specimen that is malignant to one that is benign.

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Step_15_Artefact_spec_to_spec_cont_B_1317_cropped
Section of H&E stained lung containing a piece of foreign tissue (liver) impacted into the surface at cut-up.

Take Care with Biopsy Pads

Fresh or incompletely fixed specimens are not placed between foam biopsy pads, particularly needle-core specimens (biopsy pad artifact is avoided).

Sometimes small, fresh or incompletely fixed specimens are placed between biopsy pads, put into a cassette and then fixed. This can produce a characteristic artifact.

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Step_16_Artefact_biopsy_pad_LP
The triangular spaces visible in this section result from local pressure effects caused by the cellular structure of the foam pads when applied to fresh or very briefly fixed tissue.

Choose Appropriate Cassettes

Choose appropriate cassettes for the specimen type being processed. Tissue fragments shrink during processing and, if cassette perforations are too large, fragments may escape into processing reagents or, worse still, transfer over to another specimen.

A “one size fits all” approach is used when placing specimens into cassettes.

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Step_17_One-OpenCassette-Specimen_loss_thro_holes_in_cassette
A. Some of the smaller tissue fragments seen here may escape through the holes in the cassette. This will become even more likely as the tissue shrinks during processing. B. Cassettes with fine perforations are available for small tissue fragments.

Avoid Overloading Cassettes

Cassettes are never overloaded with tissue thus allowing ready access to processing reagents and preventing distortion of specimens. If the volume of tissue is too great a second cassette is used.

Cassettes are often crammed full of tissue thus preventing access of processing reagents. Sometimes specimens are distorted in the process.

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Step_18_Over-filled_cassette
These cassettes are over-loaded. If processing goes ahead the specimens will be distorted and it is likely that the processing will be incomplete.

Clearly Label Cassettes

 Cassettes are always clearly labeled. Accurate identification of specimens is of paramount importance.

 Sometimes it is difficult to read the labels on cassettes. A bit of guess work may be required.

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Step_19_Illegible_cassette_labels
These illegible cassette labels are totally unacceptable.

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About the presenter

Geoffrey Rolls, BAppSc, FAIMS

Geoffrey Rolls is a Histology Consultant with decades of experience in the field. He is a former Senior Lecturer in histopathology in the Department of Laboratory Medicine, RMIT University in Melbourne, Australia.

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