๐Ÿ”ฌ Histology & Cytology
Lesson 01 of 31

Introduction to Histopathology

Easy โฑ 20 min read ๐Ÿ“š 35 min study ๐Ÿ—“ 2025
Course Progress0%
๐Ÿ“–

Overview

Surgical pathology includes the gross and microscopic examination of resected specimens and biopsies by histopathologists for tissue diagnosis. Several precise steps must be followed to prepare tissue in a form that can be examined under the light microscope and yield a reliable diagnosis.

This foundational lesson introduces the entire workflow โ€” from receipt of surgical specimens through to dispatch of reports โ€” and covers the critical roles of grossing, gross room design, histopathology laboratory equipment, and laboratory safety.

Subject
Histology
Difficulty
Easy
Read Time
20 min
Study Time
35 min
๐ŸŽฏ

Learning Objectives

โœ… After this lesson you will be able to
  • List the steps involved in processing surgical specimens for histopathological examination
  • Explain the after-care of specimens after diagnosis is made
  • Define grossing and describe the functions of the gross room
  • List the essential equipment and features of a gross room
  • Describe the equipment present in the histopathology laboratory
  • Identify the key laboratory hazards and apply appropriate safety measures
๐Ÿ“–

Clinical Story

๐Ÿฉบ
Every Diagnosis Starts Here

A surgeon removes a suspicious lump from a patient's breast during an operation. That specimen arrives in the histopathology laboratory in a container of formalin. Before the pathologist can ever look down a microscope, the laboratory technician must receive, label, gross, process, embed, section, and stain that tissue correctly. One error at any step can lead to a wrong diagnosis. This lesson teaches you those steps.

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Steps in Processing Surgical Specimens

1
Receipt of Specimens from OT

Specimens arrive from the operating theatre (OT) in appropriate fixative containers with completed request forms including patient identity and clinical details.

2
Grossing

Macroscopic examination of the specimen with the naked eye. The pathologist describes size, shape, colour, consistency, and selects representative sections for microscopy.

3
Tissue Processing

Selected tissue blocks are dehydrated, cleared, and infiltrated with paraffin wax using an automated tissue processor.

4
Embedding

Processed tissue blocks are embedded in molten paraffin wax and oriented correctly in moulds to form solid paraffin blocks.

5
Section Cutting

A microtome cuts sections of 3โ€“5 micrometres thickness from the paraffin block. These are floated on warm water and mounted on glass slides.

6
Staining and Labelling

Sections are stained (typically H&E), coverslipped, and labelled with patient and specimen details.

7
Dispatch to Pathologist

Completed slides are dispatched to the pathologist for microscopic examination and diagnosis.

๐Ÿ—„๏ธ

After-Care of Specimens & Records

Once the diagnosis is made and reports dispatched to surgeons, the laboratory must perform the following:

ItemStorage PeriodPurpose
Glass slidesMinimum 10 yearsFuture reference, teaching, research
Requisition formsMinimum 10 yearsMedicolegal, audit, reference
Well-preserved specimensLong-termTeaching, research, museum
Reference specimens6 months โ€“ 1 yearFuture clinical reference
Unwanted specimensDiscard after useNot useful for any of the above
๐Ÿ”ฌ
Additional Procedures in Surgical Pathology Labs

Depending on resources and requirements, a surgical pathology laboratory may also perform: Cryosections, Histochemical stains, Immunohistochemistry, Electron microscopy, and advanced techniques such as in-situ hybridisation and immunofluorescence.

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Grossing

๐Ÿ“Œ
Definition

Grossing is the process by which pathology specimens are inspected with the naked eye (macroscopic examination) to obtain diagnostic information before microscopic processing.

Points to Note Before Processing

Confirm the patient identity and the anatomical site from which the specimen was obtained. Cross-check with the request form before any processing begins.

All relevant clinical history, including the surgeon's findings, suspected diagnosis, and any prior pathology, must be reviewed before grossing.

A written record of the physical appearance of the specimen โ€” size (measured), shape, colour, consistency, surface characteristics, and any lesions identified. Only a small representative portion of a large specimen goes for microscopy, so gross examination must be performed by a skilled person.

โš ๏ธ Special Handling Rules

โ€ข Only soft tissue can be cut directly into small blocks
โ€ข Bony specimens must be decalcified before processing
โ€ข Stones and teeth require special treatment before sectioning

๐Ÿ—๏ธ

Gross Room Design & Equipment

The gross room is the dedicated space where macroscopic examination and dissection of specimens takes place.

Design Requirements

โœ…
Essential Features

The room must be large enough for all pathologists to work simultaneously. It must be well illuminated and well ventilated with an exhaust fan to remove formalin vapours.

Essential Equipment

๐Ÿ”ช
Cutting Board
Fluid must drain directly into sink
๐Ÿ—„๏ธ
Shelves for Specimen Containers
Organised by accession number
๐Ÿšฐ
Hot & Cold Water Access
Readily accessible
๐Ÿงช
Formalin (10%)
Ready access; large container
๐Ÿ”ง
Instrument Box
Forceps, scissors, probe, saw, scalpel, blades, knife, ruler
๐Ÿ“ฆ
Cassettes & Labels
For tissue sampling

Additional Items (Best Practice)

โ„๏ธ Refrigerator
๐Ÿ“ธ Photographic facility
โš–๏ธ Balance for gross specimens
๐Ÿฉป X-ray view box
๐Ÿงด Other fixatives
๐Ÿ”ฌ

Histopathology Laboratory Equipment

The histopathology laboratory must be large enough to accommodate all equipment and personnel comfortably. Core equipment includes:

โš™๏ธ
Tissue Processor
Automated dehydration, clearing, infiltration
๐ŸงŠ
Tissue Embedding Table
Paraffin wax embedding station
โœ‚๏ธ
Microtome
Cuts sections 3โ€“5 ฮผm thick
๐ŸŒก๏ธ
Tissue Warming Plate
Flattens sections before mounting
๐Ÿ’ง
Tissue Flotation Bath
Warm water bath for section floating
๐ŸŽจ
Slide Stainer
Automated or manual staining
โš ๏ธ

Laboratory Hazards & Safety Measures

โš ๏ธ Formalin Hazard

Formalin vapours are irritant to eyes and throat. Always use an exhaust fan as an outlet for vapours. Never work in a closed gross room without ventilation.

๐Ÿงค
Personal Protective Equipment (PPE)

Always wear mask, apron, eye glasses and gloves to protect against: infected material, formalin vapours, and spilt blood or body fluids.

โœ… Gross Room Safety Rules
  • Keep the grossing table clean with antiseptic solution at all times
  • All specimens must be in containers with 10% formalin and covered with a lid
  • After grossing, specimens must be stored according to their accession number
โš ๏ธ Fire Hazard

Many chemicals in the histopathology lab are inflammable โ€” wax, xylene, alcohol, acetone. Store these at a separate designated location. Minimum quantities should be kept in the laboratory. A fire extinguisher must always be available.

โš ๏ธ Electrical Equipment

Most equipment runs 24 hours, 7 days a week. Check all electrical connections before leaving the laboratory every day.

๐Ÿงค
Chemical Safety

Some chemicals are carcinogenic or harmful to skin. Always wear gloves for staining and all chemical-handling procedures.

โ“

Interactive Quiz

5 Questions
Lesson 01 โ€” Histology Quiz
5 Questionsโฑ ~5 min
Multiple Choice ยท Question 1 of 5
Slides and requisition forms in a histopathology laboratory must be preserved for at least how many years?
Multiple Choice ยท Question 2 of 5
What is GROSSING in histopathology?
True or False ยท Question 3 of 5
All specimens, including stones and teeth, can be cut directly into small blocks and processed without any special pre-treatment.
Fill in the Blank ยท Question 4 of 5
All specimens in the gross room must be stored in containers with __________ solution.
Multiple Choice ยท Question 5 of 5
Which of the following is NOT listed as essential equipment in the gross room?
๐Ÿ—‚๏ธ

Flashcards

Tap to flip
Term
What is Surgical Pathology?
๐Ÿ‘† Tap to reveal
Answer
Gross and microscopic examination of resected specimens and biopsies by histopathologists for tissue diagnosis
๐Ÿ‘† Tap to flip back
Term
What is Grossing?
๐Ÿ‘† Tap to reveal
Answer
Macroscopic (naked-eye) inspection of pathology specimens to obtain diagnostic information before microscopic examination
๐Ÿ‘† Tap to flip back
Term
How long must slides be kept?
๐Ÿ‘† Tap to reveal
Answer
At least 10 years โ€” for future reference, teaching, research and medicolegal purposes
๐Ÿ‘† Tap to flip back
Term
Why must the gross room have an exhaust fan?
๐Ÿ‘† Tap to reveal
Answer
To remove formalin vapours, which are irritant to the eyes and throat. Adequate ventilation is essential for staff safety
๐Ÿ‘† Tap to flip back
Term
Name 4 pieces of equipment in the histopathology laboratory
๐Ÿ‘† Tap to reveal
Answer
Tissue processor ยท Tissue embedding table ยท Microtome ยท Tissue flotation bath ยท Warming plate ยท Slide stainer
๐Ÿ‘† Tap to flip back
Term
What special pre-treatment do bony specimens need before processing?
๐Ÿ‘† Tap to reveal
Answer
Decalcification โ€” removal of calcium salts to soften bone so it can be sectioned on a microtome
๐Ÿ‘† Tap to flip back
๐Ÿ“‹

Case Study

๐Ÿ‘ค
Laboratory Scenario (Fictional)
Histopathology Department ยท Teaching Hospital

A surgical specimen labelled "Right mastectomy" arrives in the histopathology lab. The container is sealed with 10% formalin. The accompanying request form shows patient name, age, and clinical note: "Suspicious mass, right breast, FNAC showed malignant cells." The specimen is given accession number HP-2025-0437.

Step 1
Gross examination & description
Step 2
Representative blocks selected
Step 3
Tissue processing overnight
Step 4
Embedding, sectioning, H&E staining
  • โ†’Every specimen must have a unique accession number before any processing begins
  • โ†’Clinical details guide which blocks to sample during grossing
  • โ†’The entire 7-step process must be completed correctly before diagnosis is possible
๐Ÿ“

Quick Revision

10-Minute Review
Point 01
Surgical pathology = gross + microscopic examination of specimens and biopsies for tissue diagnosis
Point 02
7 steps: Receipt โ†’ Grossing โ†’ Processing โ†’ Embedding โ†’ Section cutting โ†’ Staining โ†’ Dispatch
Point 03
Slides and requisition forms: stored minimum 10 years
Point 04
Grossing = naked-eye macroscopic examination of specimen to obtain diagnostic information
Point 05
Bony specimens need decalcification before processing. Stones and teeth need special treatment
Point 06
Gross room must be large, well illuminated, well ventilated with exhaust fan for formalin vapours
Point 07
All specimens stored in 10% formalin in covered containers, organised by accession number
Point 08
PPE in gross room: gloves + mask + apron + eye glasses โ€” mandatory for all work
Point 09
Histo lab hazards: inflammable chemicals (xylene, alcohol, wax) โ€” store separately, fire extinguisher always present
Point 10
Electrical equipment runs 24/7 โ€” check connections before leaving the laboratory every day
๐Ÿ”‘

Key Takeaways

๐ŸŽ“ What You Have Learnt
  • Surgical pathology includes gross and microscopic examination of resected specimens and biopsies for tissue diagnosis
  • The 7 steps are: receipt, grossing, tissue processing, embedding, section cutting, staining, and dispatch
  • Slides and requisition forms must be stored for at least 10 years
  • Grossing is macroscopic inspection of specimens with the naked eye to obtain diagnostic information
  • The gross room must be large, well illuminated, well ventilated with exhaust fan, and stocked with all essential instruments
  • All specimens must be stored in 10% formalin in covered containers
  • PPE โ€” gloves, mask, apron, eye glasses โ€” must be worn in the gross room at all times
  • Inflammable chemicals must be stored separately; a fire extinguisher must always be available in the histopathology laboratory
โ˜‘๏ธ

Competency Checklist

โ˜‘๏ธ Lesson 01 โ€” Competency
0/8 complete
I can list the 7 steps in processing surgical specimens in correct order
I can define grossing and explain its purpose
I know the minimum storage period for slides and requisition forms (10 years)
I can describe the essential design features and equipment of a gross room
I know the special pre-treatment needed for bony specimens and stones/teeth
I can name the equipment present in a histopathology laboratory
I can identify the main hazards in both the gross room and histopathology lab and state the safety measures
I passed the Lesson 01 Histology quiz
Competency progress