Museum preparation in dept of pathology

1. MUSEUM
PREPARATION
MODERATOR –
DR AKASH P BHUYAN
ASST. PROFESSOR, DEPT. OF PATHOLOGY
TEZPUR MEDICAL COLLEGE.
PRESENTED BY- DR PRIYANKA GAUTAM
2ND YEAR PGT.

2. INTRODUCTION
• ALL TEACHING HOSPITALS AND COLLEGES OF PATHOLOGY HAVE MUSEUMS WHICH SERVE
MANY FUNCTIONS:
 PERMANENT EXHIBITON OF COMMON SPECIMEN FOR UNDERGRADUATE AND
POST GRADUATE TEACHING PURPOSES
 ILLUSTRATING SPECIMENS OF RARITY
 PERMANENT SOURCE OF HISTOLOGIC MATERIALS
 FOR GROSS AND MICROSCOPIC PHOTOGRAPHY

3. BASIC MUSEUM PREPARATION
1. RECEPTION
2. PREPARATION
3. FIXATION
4. RESTORATION
5. PRESERVATION
6. PRESENTATION

4. RECEPTION OF THE SPECIMEN
• ANY SPECIMEN RECEIVED IN THE MUSEUM SHOULD BE RECORDED IN THE
MUSEUM BOOK AND GIVEN A NUMBER FOLLOWED BY YEAR ( E.G 20/2023).
• THIS NUMBER WILL STAY WITH THE SPECIMEN EVEN AFTER IT IS CATALOGUED IN
ITS RESPECTIVE PLACE.
• THIS NUMBER IS WRITTEN ON TIE-ON LABEL AND STITCH TO THE SPECIMEN.
• THE RECEPTION BOOK SHOULD CONTAIN ALL NECESSARY INFORMATION ABOUT
THE SPECIMEN ( CLINICAL ,GROSS AND MICROSCOPIC FINDINGS).

5. PREPARTION OF THE SPECIMEN
• SPECIMEN SHOULD BE WASHED ONLY WITH SALINE.
• IT SHOULD NOT REMAIN IN SALINE FOR MORE THAN TWO HOURS AS AUTOLYSIS
QUICKLY SETS IN.
• COMMONEST CAUSE OF INFERIOR QUALITY OF SPECIMEN- CONTACT WITH TAP
WATER.

6. FIXATION OF THE SPECIMEN
• OBJECTIVE- TO PRESERVE CELLS AND TISSUE CONSTITUTENTS AS CLOSE TO
LIFE- LIKE STATE AS POSSIBLE.
• FIXATION ARRESTS AUTOLYSIS AND BACTERIAL DECOMPOSITION AND
STABILISES THE CELLULAR AND TISSUE CONSTITUTENTS.
• FIXATIVE USED – KAISERLING TECHNIQUE ( FORMALIN FIXATIVE TECHNIQUE)
AND ITS MODIFICATIONS.

7. FIXATION CONTD..
• KAISERLING RECOMMENDED THAT THE INITIAL FIXATION SHOULD BE NEUTRAL
FORMALIN (KI) SOLUTION AND THAN TRANSFER TO A FINAL PRESERVING
GLYCERINE SOLUTION (KIII) FOR LONG TERM DISPLAY.
• COLOR PRESERVATION IS ALSO MAINTAINED WITH THESE SOLUTIONS.

8. PRINCIPLES OF FIXATION
• SPECIMEN CONTAINING BILE OR STAINED BY BILE MUST BE FIXED ANSD STORED
APART FROM OTHERS
• SPECIMEN UNDERGOING FIXATION MUST NOT TOUCH OTHER SPECIMEN OR THE
SIDES 0F JAR.
• THEY SHOULD BE SUSPENED BY LINEN THREAD.
• SPECIMEN OF FLAT TISSUE LIKE STOMACH , INTESTINE FIXED TO CORK BOARD
AND LEFT IN FORMALIN TO AVOID IRREGULAR FIXATION AND CRUMPLING.

9. PRINCIPLE OF FIXATION CONTD..
• CYSTIC CAVITIES-
IF UNOPENED: INJECT WITH FIXATIVE.
IF OPENED: PACK WITH COTTON WOOL.
• SOLID VISCERA-
FIXED BY VASCULAR INJECTION (BRAIN THROUGH BASILAR ARTERY)

10. FIXATION TECHNIQUES
• MOST WIDELY USED IS KAISERLING TECHNIQUE.
• THE ORIGINAL TECHNIQUE USED 3 SOLUTIONS-
1) FOR FIXING
2) FOR RESTORING COLOR
3) FOR MOUNTING FLUID.

11. • KAISERLING I SOLUTION-
FORMALIN 1L
POTASSIUM ACETATE 45GM
POTASSIUM NITRATE 25GM
DISTILLED WATER UPTO 1L
• SPECIMEN IS STORED IN THIS SOLUTION FOR 1 MONTH DEPENDING ON THE SIZE
OF THE SPECIMEN.

12. RESTORATION OF SPECIMEN
• THE RECOMMENDED METHOD IS KAISERLING II METHOD ( 80-95% ETHYL
ALCOHOL).
• STEPS-
a. REMOVE THE SPECIMEN
b. WASH IT IN RUNNING WATER
c. TRANSFER TO ETHYL ALCOHOL SOLUTION FOR 10 MIN TO 1 HOUR
DEPENDING ON THE SIZE OF SPECIMEN.
d. SPECIMEN IS THEN OBSERVED FOR COLOR CHANGE.
e. SPECIMEN IS READY FOR PRESERVATION.

13. COLOR RESTORATION
• REJUVENATOR SOLUTION RESTORES THE COLOR.
• REJUVENATOR SOLUTION
1. PYRIDINE 100ML
2. SODIUM HYDROSULPHITE 100GM
3. DISTILLED WATER 4 LTRS

14. HOLLOW VISCERA
• CUT HOLLOW ORGANS SHOULD BE PADDED OUT WITH
COTTON WOOL; UNCUT ORGANS CAN BE PRESSURE
INFLATED.
BLADDER & PELVICALYCEAL SYSTEM- THROUGH
URETHRA.
LUNG- THROUGH TRACHEA.
CYSTS- DIRECT INJECTION.
• FIXATIVE CAN BE INJECTED WITH HIGGINSON SYRINGE.

15. SPECIMEN OF HEART
• CUT SPECIMEN- PAD OUT ALL CAVITIES AND
MAJOR VESSELS WITH COTTON WOOL BEFORE
FIXATION TO MAINTAIN THE ORIGINAL SHAPE.
• FRESH AND UNCUT HEART- PLACE IN A
LARGE CONTAINER OF FIXATIVE AND ALSO
INJECT THROUGH CORONARY OSTIA.

16. SPECIMEN OF BRAIN
• FIX THE BRAIN BEFORE CUTTING.
• PERFUSE THE BRAIN THROUGH THE
BASILAR AND CEREBRAL ARTERIES AT
ITS BASE AND IT SHOULD THEN BE
SUSPENDED BY THE BASILAR ARTERY
AT LEAST FOR A WEEK.

17. SPECIMEN OF BRAIN CONT..
• AFTER FIXATION, PH SHOULD BE DETERMINED.
• IF PH > 6.5, SPECIMEN SHOULD BE PLACED DIRECTLY IN KAISERLING III
SOLUTION.
• IF PH < 6.5, SPECIMEN IS PLACED IN KAISERLING II SOLUTION.

18. FACTORS AFFECTING FIXATION
• BUFFERING
• PENETRATION
• VOLUME
• TEMPERATURE
• CONCENTRATION

19. BUFFERING
• FIXATIVES ARE GENERALLY BUFFERED TO A PH BETWEEN 6-8.
• ACIDIC PH FAVORS FORMATION OF FORMALIN-HEME PIGMENT THAT APPEARS AS
BLACK, POLARIZED DEPOSITS IN TISSUE.
• COMMON BUFFERS USED- PHOSPHATE, BICARBONATE .
• COMMERCIAL FORMALIN IS BUFFERED WITH PHOSPHATE AT A PH OF 7.

20. PENETRATION
• IT DEPENDS UPON THE DIFFUSABILITY OF EACH INDIVIDUAL FIXATIVE.
• FORMALIN AND ALCOHOL PENETRATE THE BEST.

21. VOLUME
• STANDARD RATIO OF FIXATIVE TO TISSUE VOLUME IS 10:1.
• CAUSE OF POOR TISSUE PRESERVATION- USE OF LOWER VOLUME
OF FIXATION FLUIDS FOR LARGER SPECIMEN.

22. TEMPERATURE
• INCREASING THE TEMPERATURE WILL INCREASE THE SPEED OF
FIXATION.
• HOT FORMALIN WILL FIX TISSUES FASTER.

23. CONCENTRATION
 CONCENTRATION OF THE FIXATIVES AFFECT THE RATE OF FIXATION AND
PENETRATION OF FIXATIVE INTO THE SAMPLE.
 HIGH CONCENTRATION- CAUSE HARDENING OF THE TISSUE.
 LOW CONCENTRATION- CAUSE EXHAUSTION OF THE FIXATIVE BEFORE THE
PROCESS IS COMPLETE.

24. • FIXATION:
a) CONFERS CHEMICAL STABILITY ON THE TISSUE.
b) HARDENS THE TISSUE.
c) STOPS ENZYMATIC AUTOLYSIS AND BACTERIAL
PUTREFACTION.

25. PRESERVATION
• THE SOLUTION RECOMMENDED FOR THIS STEP IS KAISERLING III.
• IT IS BASED ON GLYCERINE SOLUTION.
KAISERLING III SOLUTION:
POTASSIUM ACETATE 1416GM
GLYCERINE 4L
DISTILLED WATER MAKE UPTO 10L.
• THYMOL CRYSTALS ADDED TO PREVENT MOULDS.

26. • LEAVE SOLUTION TO STAND FOR 2-3 DAYS BEFORE USING TO ENSURE
PROPER MIXING OF SOLUTION.
• STABILIZER:
 1% PYRIDINE IS USED.
 ACTS AS PERMANENT FIXATIVE.
 REPLACE FREQUENTLY TO RESTORE COLOR OF THE SPECIMEN.

27. PRESENTATION
• MUSEUM SPECIMEN ARE GENERALLY MOUNTED
IN GLASS JAR OR PERSPEX BOXES USING CENTER
PLATES OR GLASS RODS TO WHICH THE
SPECIMEN IS FIXED OR SUTURED.
• ALSO POLYETHYLENE TETRAPHTHALATE (PET,
PETE, OR POLYESTER) CAN BE USED.
PET SHEETS CAN BE EASILY CUT
, SUTURED TO THE SPECIMEN ( MULTIPLE IF
NEEDED) AND DOES NOT EASILY HINDER THE
DISPLAY.
FOR SMALLER JAR HINGE LIKE
PREPARATION OF THE PLASTIC BOTTLE GIVES A
STABLE SUPPORT TO THE SPECIMEN

28. FOR SMALL AND FRAGILE TISSUE THAT MIGHT TEAR ON
APPLICATION OF SUTURE CAN BE SIMPLY STUCK TO THE
COMERCIALLY AVAILABLE MICROSLIDE.
SLIDES CAN BE JOINED TOGETHER USING DPX MOUNTANT
TO GET REQUIRED SHAPES AND SIZES SUITABLE FOR THE
SPECIMEN TO BE DISPLAYED AGAINST.

29. • PERSPEX BOXES ARE USED COMMONLY.
• POSITION IN WHICH THEY ARE TO BE MOUNTED SHOULD BE ANATOMICALLY
CORRECT.
• ½ INCH BOTTOM CLEARANCE IS ALLOWED FOR A LABEL TO BE FITTED WITHOUT
OBSCURING PART OF THE SPECIMEN.
• TO SUPPORT THE SPECIMEN WITHIN JAR, IT IS ATTACHED TO THE SPECIMEN
PLATE OR GLASS RODS

30. • SPECIMEN IS ARRANGED IN THE DESIRED POSITION AND CROSSES ARE MADE ON
THE PLATE WHERE STITCHES ARE TO BE PLACED TO HOLD THEM IN POSITION.
• STITCHES MUST NOT BE PLACED THROUGH PATHOLOGICAL LESIONS.
• SPECIMENS ARE TIED WITH NYLON OR LINEN THREADS AND PUT INTO THE BOX.

31. SPECIAL METHODS
• MACERATION
USED TO DEMONSTRATE BONY LESION .
ENABLES PRESERVATION OF EVEN THE FINEST BONY SPICULES.
• STEPS-
TRIM OFF THE EXCESS SOFT TISSUE
BONE IS EMERGED IN CHLOROFORM FOR 3 TO 4 HOURS TO REMOVE FAT
DRIED IN INCUBATOR AND BLEACHED IN HYDROGEN PEROXIDE
• MACERATED BONES ARE MOUNTED DRY AND FIXED WITH NYLON WIRES.

32. AMYLOID
IODINE TECHNIQUE
• FORMALIN FIXED TISSUE IS PLACED IN
LUGOL’S IODINE
• 1% SULPHURIC ACID IS ADDED AND
KEPT FOR 1-2 HRS
• WASH IN RUNNING WATER
• MOUNT IN LIQUID PARAFFIN
CONGO RED TECHNIQUE
• AFTER FIXATION SPECIMEN IS IMMERSED IN
1 % CONGO RED FOR 1 HOUR
• TRANSFER TO SATURATED SOLUTION OF
LITHIUM CARBONATE FOR 2 MINS
• DIFFERENTIATED IN 80% ALCOHOL
• NORMAL ARTERIES AND VEIN RETAIN THE
COLOUR
• SPECIMEN MOUNTED IN KAISERLING
SOLUTION III

33. • MUSEUM SPECIMEN SHOULD BE CLEARLY LABELLED AND
A SYSTEM OF CATALOGUING SHOULD BE EMPLOYED
WHICH ALLOW EASY ACCESS.