This document provides guidelines for handling radical prostatectomy specimens to ensure accurate pathological assessment. It discusses transporting specimens in formalin on ice, weighing, measuring, harvesting fresh tissue, formalin fixation, inking, slicing, sampling seminal vesicles, and total versus partial embedding. Standardized handling is important because accurate reporting guides adjuvant therapy for high-risk prostate cancer patients.

1. Handling Of Radical
Prostatectomy
Specimens
Adapted from a review :-
Handling of radical prostatectomy specimen.
Histopathology 2012; 60, 118-124
Dept of Oncology-Pathology, Karolinska Institute, stock-Holm, Sweden.
Thanks to Dr Farzana Habib, our PG Trainee

2. BRIEF SUMMARY
 This is a review article by Lars Egevad
 Accurate reporting of radical
prostatectomy specimens becoming more
important
 Adjuvant therapy given to pts with
pathological findings indicating a high risk
of disease recurrence
 Handling must therefore be standardized
 Guidelines for gross description, bio
banking of fresh tissue, fixation and cutting

3. INTRODUCTION
 Handling of radical prostatectomy specimens
is a challenging task for the pathologist
because: prostate undergoes faster autolysis
than most other organs
 Prostate cancer difficult to identify with naked
eye
 Tumors smaller yet more multifocal
 Ca very heterogeneous both morphologically
and genetically
 Thus these specimens need to be handled
with great care and according to standardized

4. TRANSPORTATION OF
SPECIMEN
 Buffered 4% formaldehyde solution
 Impairs harvesting of fresh tissue
 Unfixed material needed for hormonal assays and
molecular analyses
 Volume of formalin 10 times the specimen volume
(500ml)
 Proteolytic enzymes in prostatic secretions make it
more sensitive to autolysis (transported rapidly,
kept on ice)

5. WEIGHING AND MEASURING
 Seminal vesicles and vas deferens
removed prior to weighing and measuring
 Three dimensions

6. HARVESTING OF FRESH TISSUE
 For research purpose, 55.4% academic and
7.2% non academic
 Several methods have been described
 Core biopsies
 Shave sections or Punch biopsies
 Cytological method
 Freezing of entire section
 Freezing of entire slice (liquid nitrogen)
 Steu et al recommended precooled isopentane
or liquid nitrogen rather than CO2 snow
 Final cutting of the prostate after formalin
fixation

7. FORMALIN FIXATION
 2.4mm per 24h
 < 10mm simple diffusion, > more than 48h
 Large prostate 6-7cm, several days
 Autolytic artefacts impair h/p dx.
 Features of prostate autolysis include retraction
of glandular epithelium from the stroma with
collapse of glandular units
 Several methods
 Formalin injection using 20ml syringe
 Aquarium pump or magnetic stirrer
 Microwaving for 1-2 min

8. INKING AND SLICING
 Evaluation of surgical margins
 Positive surgical margin increases
recurrence
 By definition , cancer glands must reach
ink for the margin to be considered
positive
 Minimum 2 different colors for right and left
 Adhesion of ink improved by 5% acetic
acid
 Apex cut by a modified cone method

9. Sampling with routine sections.
Each transverse slice of prostate
gland (A) is further cut into two
halves (B) or four quadrants (C) to
accommodate the size of
conventional cassettes (D).
The entire prostate was carefully
inked by painting the surface with
different colours of ink to ensure
proper orientation and margin
identification. The right lobe was
painted yellow, and the left lobe
black.

10. SEMINAL VESICLES
 Invasion associated with poor prognosis
 Defined as Tumor cells invading the
muscular coat of extra prostatic portion of
seminal vesicles
 Embedding of entire seminal vesicles not
mandatory, basal portion and transition to
prostate examined (min requirement)
 Embedding of vasdeferens margin not
obligatory

11. FIGURE A, THE PROSTATE SHOULD BE WEIGHED AND MEASURED AFTER THE SEMINAL VESICLES HAVE
BEEN REMOVED. B, HARVESTING OF FRESH TISSUE BY
SHAVING FROM THE CUT SURFACE. C, HARVESTING OF FRESH TISSUE BY SPLITTING A SLICE OF THE
PROSTATE IN SEGMENTS. D, INJECTION OF FORMALIN FOR ENHANCED
FIXATION. E, THE PROSTATE MUST BE MOUNTED AND THE CAPSULE PINNED DOWN IF FRESH TISSUE HAS
BEEN HARVESTED. F, THE PROSTATE IS INKED WITH AT
LEAST TWO COLOURS AFTER FIXATION.

12. TOTAL v/s PARTIAL
EMBEDDING
 Prostate cancer not visible at gross
 Safest method is that entire prostate is
submitted
 Sehdev et al compared 10 protocols for subtotal
embedding, every post. Quadrant and mid ant.
Section on each side, saved 7 standard blocks
 Subtotal embedding can decrease the work
load but additional cutting and new blocks
delays the report and adds further to the work
load

13. WHOLE-MOUNT v/s
STANDARD SECTIONS
 Whole-mount sections
• More difficult to make
• More expensive
• Difficult to perform IHC
• Don’t fit into standard slide holders
 Give a better overview and identification
of multiple tumor foci, lab techs find them
less time consuming
Figure . A representative whole-mount section of prostate gland.

14. Table 1. Obligatory procedures for handling of
radical prostatectomy specimens
 Removal of seminal vesicles before weighing of
prostate
 Recording of weight of prostate
 Recording of three diameters of prostate
 Inking of prostate with at least two colours
 Slicing after full fixation
 Modified cone method, apex
 Modified cone method, base
 If partial embedding is used, method should be
documented
 Embedding of section through the base of the
seminal vesicle

15. FUTURE PERSPECTIVES
 Clinical management
 Adjuvant treatment
 In the near future, h/p examination of the
specimen will have critical importance for
pt care
 Fresh tumor tissue for research purpose
 Therefore it is important that standardized
protocols are developed for the handling of
radical prostatectomy specimens