How to collect and send specimen for histopathology in correct container is discussed here.

1. WELCOME

2. Presented By
Dr. Mansurul Islam(Anik)
Asst. Reg. SU-II
Department of Surgery, ShSMCH
Sending Specimen For
Histopathology

3. Beginning of Microscopy
• In the past, tissue diagnosis was restricted to macroscopic
examination of autopsy material and of a limited range of
surgical specimens.
• In the nineteenth century, microscopic examination of human tissue from
autopsies and surgical procedures was introduced.
• Tissue analysis is now an integral and routine element of clinical
management.

4. Reasons for tissue analysis
• To make a new diagnosis
• To confirm a suspected or established clinical diagnosis
• To exclude additional diagnoses
e.g. An appendix removed for appendicitis could contain an incidental
carcinoid tumour

5. Reasons for tissue analysis(Cont.)
• To assist with prognosis
• To stage tumours
• To help select therapy and plan management
e.g. A cancer resection specimen confirms the diagnosis and also
provides information about features such as tumour stage, vascular invasion
and resection margin involvement, which in turn predict clinical outcome
and help determine postoperative treatment.

6. Reasons for tissue analysis(Cont.)
• To assess response to treatment
e.g. Pathologists are to assess the degree of tumour regression after
neoadjuvant therapy
• Audit
e.g. Surgeons and Radiologists can audit their performance.

7. Histology
Formalin-fixed tissue
 Biopsy
Mucosal
Punch
Needle (core)
Curettings
Excision
Resection
Fresh tissue (usually for frozen section)

8. Common types of tissue sample(Cont.)
Cytology
• Cervical
• Washings, brushings, scrapes
• Fine-needle aspirate (FNA)
• Fluids/sputum

9. Histology
Biopsy:
Taken for diagnosis and assessment rather than treatment.
Excision biopsy:
Serves as both a diagnostic biopsy and a limited resection.
Resection:
Done to remove a lesion (e.g. a tumour) but have other uses including
diagnosis, staging, and determination of further management.

10. How to send specimen for histopathology?
Care should be taken-
• During and after the biopsy surgery
• During fixation and transport
• Important information to be communicated to pathologists

11. Care During and After the Biopsy Surgery
• Avoid contamination of the tissue with extraneous material, particularly
tissue from another patient.
• Avoid Artifacts-
 Injection Artifacts –
 Causes separation of connective tissue bundles.
 Infiltration of anesthetic agents directly into the lesion should be avoided.
 Suction Artifacts-
 Formation of large often pleomorphic connective tissue vacuoles
resembling traumatized adipose tissue.

12.  Cautery Artifacts-
 Tissue shows a coagulated and torn appearance.
 Only the cutting and not the coagulation electrode should be used.
Forceps Artifacts-
 tears and compression of the surrounding tissue.
 Surface epithelium may be forced through the connective tissue producing
small “pseudocysts”.
 Using small atraumatic forceps or a suture placed to the edge may be used
as a substitute.

13. Forceps Artifact Injection Artifact

14. Care During Fixation and Transport
 Fixation-It is a complex series of chemical events which brings about
changes in the various chemical constituents of cell, however the cell
morphology and structural detail is preserved.

15. Advantages of Fixation
1. To preserve the tissue as much possible.
2. To prevent postmortem changes like autolysis and putrefaction.
3. Preservation of chemical compounds and microanatomic constituents so that
further histochemistry is possible.
4. Hardening : the hardening effect of fixatives allows thin sectioning.
5. Effects of staining : certain fixatives like formaldehyde intensifies the staining
character of tissue.
6. Kills microbes.

16. Fixation
• Done as soon as possible.
• Effective and appropriate fixative.
• Routine surgical specimens in 10% buffered formalin.
• Delay in fixation and inadequate fixation alters the staining quality of
the cells.
• For routine histopathology, fixation should be carried out at room
temperature.

17. Fixation (Cont.)
• Hollow specimen and cystic cavities are either opened or else fixed
simultaneously from the outside and inside. Hollow specimen cavity is
filled with formalin by syringe or catheter or packed with gauge or
cotton impregnated with formalin. Cystic lesions are injected with
formalin after the original fluid has been removed. Multilocular cysts
require individual injections.

18. Container
• Specimens should be in a container that can be sealed
• Container should not leak
• It should be large enough to accommodate the specimen and filled with
enough formalin to completely cover the specimen.
• The specimen should be able to float freely in the container for adequate
fixation.
• The container should have an opening large enough so that the tissue can
be removed easily after it has been hardened by the fixation.

22. Container(Cont.)
• Must be properly labelled.
• Any risk of contamination by transmissible infection, e.g.hepatitis B virus,
must be minimised by the use of warning labels, especially when fresh tissue is
being submitted.
• Formalin itself is toxic to the eyes and skin. Formalin toxic label must be added.

23. Surgical Pathology Specimens Requiring Special
Studies:
• CHROMOSOME STUDIES: Tissue that requires chromosome studies
should be sent fresh not in formalin. This includes products of
conception, fetal tissue or placental tissue.
• CULTURES: Any tissue requiring cultures should be sent fresh and not
in formalin.
• FROZEN SECTION: Surgical pathology specimens that need frozen
section should be fresh and not in formalin.

24. • MUSCLE/NERVE BIOPSIES: Muscle/ Nerve biopsies are sent fresh and not
in formalin. A few drops of saline can be added if the tissue is at risk to dry
out.
• SKIN BIOPSIES FOR IMMUNOFLOURESCENCE: Special skin biopsies that
require immunofluorescence studies should have one piece sent fresh with
no formalin. A separate piece is needed for histology and is fixed in
formalin.

25. Transport
• It should be done in appropriate sealed containers and should be as
soon as possible.
• If delay is unavoidable, it is better to communicate to pathologist and
to ascertain with him the type of tissue and proper fixative required.

26. Care in delivering information to Pathologists
• Always tissue has to be sent with a proper requisition form, given an
identification number.
• All relevant details like site, size, shape, dimensions etc. of specimen
must be mentioned.
• Dimensions as seen clinically and after tissue has been excised has to
be mentioned clearly as tissue may swell up or shrink in fixative.

27. • Some specimen needs proper marking with sutures.
• A brief history with suspected diagnosis must be added.
• With historical background, physical findings and precise orientation
of anatomic relations, the pathologist can block the tissue in the
plane that will give meaningful sections.

28. Resected Specimen of Brest

29. Resected Specimen of Stomach

30. • The urgency of the report should be mentioned.
• Contact details of the surgeon must be mentioned so that pathologist can
consult regarding any query.
• In case of review of any report the cause of review and previous report
must be delivered.
• The surgeon may already know the microscopic diagnosis and is now
interested in information such as extent of lesion, invasion of neighboring
structures, presence of tumour at surgical margins, vascular invasion and
lymph node metastasis, these things must be mentioned.

31. If we expect Proper Histopathological report from Pathologists-
Proper Biopsy procedure
Proper Fixation
Proper Labelling
Proper Requisition
Proper Transport must be ensured.