HISTOPATH LAB (MODULE 2) Requirement to send specimen

RECEIVING, ACCESSIONING AND GROSS  Requisition form/submission form

EXAMINATION OF TISSUE SAMPLES  Container
 Fixative
Specimens received for histological examination
may come from a number of different sources. Specimen handling and transport to laboratory
They range from very large specimens or whole
When sending large specimens or small biopsy
organs to tiny fragments of tissue. Specimens
samples:
are usually received in fixative (preservative)
but sometimes arrive fresh and must be  The specimen should be collected into a
immediately fixed. Before specimens are wide-mouthed container with a well-
accepted by a laboratory the identification fitting lid to prevent leakage of the
(labelling) and accompanying documentation will fixative containing an adequate amount
be carefully checked, all details recorded and of 10% formol saline (formalin) to
“specimen tracking” commenced. It is vital that completely submerge the specimen.
patient or research specimens are properly  For large specimens, the container should
identified and the risk of inaccuracies be larger than the specimen, preferably a
minimized.  bucket.
 Specimens should be transported to the
The following are some of the specimen-types
laboratory as soon as possible to prevent
commonly received in a histopathology lab:
decay of samples for proper fixation
 Excision specimens (surgical biopsies) - procedures to be carried out.
where whole organs or affected areas are  Specimen should be submitted in the
removed at operation laboratory with the properly filled-up
 Incisional biopsy specimens - where tissue request form
is removed for diagnosis from within an For frozen sections:
affected area
 Punch biopsies - where punches are used  The specimen should be sent in the fresh
to remove a small piece of suspicious state, without any fixative.
tissue for examination (often from the  Specimens should be submitted in the
skin) laboratory with the properly filled up
 Shave biopsies - where small fragments of request form
tissue are “shaved” from a surface
Specimen identification and labeling
(usually skin)
 Curettings - where tissue is removed in A properly filled-up surgical pathology request
small pieces from the lining of the uterus form must accompany the tissue specimen
or cervix received in the laboratory.
 Core biopsies - where a small tissue
 Patient information – middle name,
sample is removed using a special needle
birthday, hospital number and other
sometimes through the skin
demographic data
(percutaneously).
 Patient’s history
Specimen Receiving and Preparation  Physical, laboratory and imaging findings
 Patient’s diagnosis – pre-operative or
Tissue for study can be obtained from post-operative diagnosis
 Biopsy – sample of tissue taken from the  Description of the site of origin
body in order to examine it more closely
using a microscope in the laboratory
o A doctor/surgeon will make a cut
or incision of the patient’s body
part in order to remove a sample
of tissue which we call biopsy.
 Autopsy – examination of the body of a
dead person. It may be restricted to a
specific organ or region of the body.
 - Once the sample is accepted then
proceed to accessioning procedure or
assign a number that will identify the
specimen for a particular patient
Gross examination
- Grossing is an art
- A knowledge of what needs be taken for
microscopic study is crucial or important
for the final diagnosis
- Grossing involves:
 Accurate naked eye description of
intact specimen (specimen
identification)
 Identify all the anatomical
structures present
 Orientation markers should be
identified
 Measurements: length, width,
height
 Weight especially parenchymatous
Specimen identification and labeling organ
Specimen container:  Examine the external surface
 Cut all the organs at intervals of 1
 Plastic or glass jar – with well fitting lid cm thickness (bread-loafing style)
or cover, make sure that the jar is bigger  Describe cut surface, identify
than the specimen pathologic process
 Label matching requisition slip  If suspected lesion is present,
 Registration no. measure, describe with color and
 Full name consistency
 Age, sex  Surgical margins
 Ward no., bed no.  Histological sections
 Site and side – where the tissue sample or
biopsy is taken Grossing room
 Multiple specimens from same patient  Large room, well illuminated and
mark as A, B, C, D etc. properly ventilated
 Signature of doctor with date  Cutting board places inside a metal box
Accessioning procedure designed in such a fashion that all the
fluids flow directly into the sink
- The specimens are accessioned by giving  Shelves for specimen container
them a number that will identify each  Ready access to a sink with hot and cold
specimen for each patient. water
- Most hospitals label specimen using  Ready access to formalin
accession number instead of patient’s  Box of instruments, box with cassettes,
name labels
- Unlabeled specimens are absolutely  Large formalin container, photographic
unacceptable for accessioning procedure facilities
- Inconsistencies between the specimen  Large table with sink for large specimens
and the request form must be returned to  Central table for multiple purpose
the operating room for correction by the
staff concerned.
- Incomplete specimens are likewise
unacceptable
- Accessioning procedure is done right
after inspecting samples
 LIMS (Laboratory Information
Management System)
 Data entered incorrectly in LIMS

- During tissue dissection/grossing

 Request form and specimen match
not performed
 Specimen containers wrongly
numbered
 Cassettes labelled with different
Inking accession number to specimen
 Sub-numbering of cassettes
- The use of an assortment of ink colors is
incorrect
useful in the following ways:
 Tissue lost or contaminated during
 For resection margins
section
 For orientation of a specimen
 Tissue inadequately fixed or
 To reduce identification error
decalcified
when multiple sampling is
required from the same tissue ALWAYS REMEMBER!
 Embedding instructions
 Histopathological examination is used to
 Identify cut surface
provide diagnostic information that is
Criteria for rejection of gross specimen important for timely diagnosis to
determine treatment plan
- Reject the specimen when:
 Loss of specimen or specimen switching is
 Discrepancies between the
tragic both for patient and pathologist
requisition and specimen labels
 Handle all specimens with care and
 Specimen with no labels
respect
 Mislabeled specimen
 Fresh tissue is extremely fragile and
 Absent clinical data or history
subject to autolysis. Therefore, handle
 Inappropriately identified
all specimens quickly and correctly
specimens
Common errors during receipt of samples and
Specimen reception (video)
gross examination
In microbiology department
- During receipt of sample
 No tissue received in specimen Segregated by specimen type:
container
 Urine
 Specimen received without
 Blood cultures
formalin
 Fluids
 Tissue samples received in
 Tissues
wrongly sized containers
 Swabs
 Number of samples not consistent
with the request form Microbiology request form:
 Incomplete or incorrectly labelled
request form or specimen  Patient hospital number
 Labelling of sample and request  Surname
form not matching  First name
 Date of birth
- During data entry  Gender
 Sample and request form  Specimen type
allocated wrong number  Test requested
 Sample and request form given  Consultant who requested test
different laboratory numbers  Location where sample was taken
(accession numbers)  Time
 Sample and request form  Date
allocated to wrong patient on
Before opening the sample, check the integrity picture of what it is to be done in the reception
first. Like for example, if it’s not broken or area upon receiving the specimen.
anything.
Gross examination by pathologist (video 2)
Sample inspection
Gross Examination of Whole Prostatectomy
 Clean with disinfectant Specimens
 Minimum identification criteria (check if
 Patient identification (verify patient’s
it corresponds correctly)
name on the container)
o Hospital number
 Specimen orientation (e.g., seminal
o Full name
vesicle)
o Date of birth
o Seminal vesicles are posterior
 Check the sample (if it’s correct with
o Superior right side is inked green
what you receive)
o Left side is inked blue
 Sample mismatch (if the name doesn’t
o Whole prostate is inked black
correspond with the name on the form or
when you received a totally unlabeled o Black ink is more permanent when
specimen, then: adhering to tissues assuring better
o Reject easily repeatable samples identification of surgical margins
and request a retake and sealing the previous green
o If not repeatable, call medic to and blue inks.
o Fixing the ink by spraying the
confirm identity of sample
specimen with acetic acid 20%
Booking in system  Begin sectioning specimen
o The seminal vesicles are sectioned
 Lab number
at their intersection with the
 Hospital number
prostate
 Patient’s surname
o Measuring
 First name
 Sex  Left and right
 Date of birth measurement
 Location  Height
- If the patient wasn’t found in the system,  Length
so new entry will be created. All the  Weight
information came across and then put  Volume measurement by
technical details: doing water displacement
 Additional information  Insert probe through
 Test which was requested urethra helps to orient the
 Specimen type axis of the prostate
 Time of collection  Sectioning procedure at
 Received date 4mm slices from apex to
- Saving these data means the quest is base
booked and this request is new to specific  Embedding piece through
lab number which is used to record bread-loaf style (two little
sample request. sections)
 Make sections with wide
Providing result movements of the blade to
avoid crinkling of the cut
 Results are only given to authorized
surface
personnel (e.g, GP’s, ward staff)
 Section the seminal vesicle
**please take note that this video presented  Labeling cassettes and submitting the
microbiologic samples but notice that protocols sections
for specimen receipt are almost the same except o Crinoline bags
that samples delivered in the histopathology
**This video shows how the grossing of a tissue
laboratory are tissues that are soaked in
specimen is done from patient identification,
fixative. This video aims to give you an actual
inking, cutting (bread-loafing style), describing,
measuring, labeling, and putting the tissue into
tissue cassettes in preparation for processing.