Upload
gulab-chand-tiwari-tiwari
View
126
Download
3
Embed Size (px)
DESCRIPTION
COMPARATIVE STUDY OF ER/PR/Cerbb2 USING FAA FIXATIVE AND 10% BUFFER FORMALINMr. G. P. Tiwari Department of Pathology Tata Memorial HospitalIntroduction ER/PR/Cerb2are the most important prognostic and predictive factors in breast carcinomas. demonstration of these biomarkers remains the main stay of assessment. it is affected by many technical factors especially fixation. IHC HoweverIntroduction ctd FAAfixative or AFA is an alcoholic based fixative with a combination of 3 c
Citation preview
COMPARATIVE STUDY OF COMPARATIVE STUDY OF ER/PR/Cerbb2 USING FAA ER/PR/Cerbb2 USING FAA FIXATIVE AND 10% BUFFER FIXATIVE AND 10% BUFFER FORMALINFORMALIN
Mr. G. P. TiwariDepartment of Pathology
Tata Memorial Hospital
IntroductionIntroduction
ER/PR/Cerb2 are the most important prognostic and predictive factors in breast carcinomas.
IHC demonstration of these biomarkers remains the main stay of assessment.
However it is affected by many technical factors especially fixation.
Introduction ctd Introduction ctd
FAA fixative or AFA is an alcoholic based fixative with a combination of 3 chemicals formaldehyde, alcohol, acetic acid. It was first described by William Davidson and William Hartmann in the year 1953.
Most European countries use alcoholic formalin to fix breast cancer tissues for better preservation of ER receptor.
QuestionsQuestions
Is FAA cost effective?Should FAA be advocated for all
round use?- as many labs may not have the resources.
AIMS & OBJECTIVESAIMS & OBJECTIVES
To evaluate if FAA helps in improving the results of ER/PR/cerb2 in breast cancer.
To compare the results with buffered formalin.
INGREDIENTS OF ORIGIONAL INGREDIENTS OF ORIGIONAL FAAFAA
ETHANOL(95-100%): 85.0 ML
FORMALIN(37-40%): 10.0 ML
GLACIAL ACETIC ACID: 5.0 ML
If the concentration of the acetic acid is 5%, it brings swelling in the tissue and Slides becomes hazy and patchy staining with IHC was observed but 0.5% acetic acid concentration is sufficient to over come this problem.
MODIFIED FAA SOLUTIONMODIFIED FAA SOLUTION
ETHANOL (95 – 100 %) :- 85.0 ML
BUFFERED FORMALIN(37-40%) :-10 ml
GLACIAL ACETIC ACID :- 0.5 ML
The amount of acetic acid or formalin may be varied.
Methanol or isopropyl alcohol may be used in stead of ethanol
Acetone, methanol, ethanol, isopropanol
Displace water molecules from pertinacious materials and extract lipids from the tissue
CHEMISTRY OF FAA FIXATIVECHEMISTRY OF FAA FIXATIVE
Acetic acid
Coagulates nucleic acid and enhances the rate of penetration of formalin in the
tissue
The tissue is fixed rapidly as compared to 10% buffered formalin
CHEMISTRY OF FAA FIXATIVECHEMISTRY OF FAA FIXATIVE
ADVANTAGES OF FAAADVANTAGES OF FAA
Inhibits formation of formalin pigment Rapid fixative(5mm in 4 hrs.) LESS-toxic Less expensive Easy to prepare Osmotic pressure is well maintained It permits all most all type of staining and
immunohistochemistry stains Turn around time is improved for 2 days
TECHNICAL CONSIDERATIONSTECHNICAL CONSIDERATIONS
The mixture can be kept indefinitely but its chemical composition changes considerably with storage..
The concentration of ethanol and formaldehyde decline, so the fixative property also declines, hence must be prepared fresh everyday.
ROUTINE PROCESSING SCHEDULE ROUTINE PROCESSING SCHEDULE FOR BREAST TISSUESFOR BREAST TISSUES
Fixation- 48 hours Dehydration- 06 hours Clearing- 06 hours Impregnation- 06 hours Total Processing Time-22-23 hours. Turn Around Time-4 days for slide
submission, 6 days after ER,PR
Material and MethodMaterial and Method
25 cases of primary operable breast cancer tissues who had not received prior chemotherapy were evaluated.
Two sections from tumour as similar as possible were taken. One was fixed in buffered formalin and the other in FAA.
The tissue sections were coded blindly unknown to the pathologist for identification as to what was the nature of fixation.
RESULTSRESULTS
H & E – Cellular swelling but less shrinkage
Out of 25 samples 10 cases are negative 13 cases are positive 2 cases are without tumor
Marker FAA BF
ER+ PR + 8++ 8
ER+ PR - 2 2
ER- PR + 1 0
ER- PR - 9 10
Cerb2 + 0 1*
Cerb2 - 20 19
Two cases were without tumor
*On review false reported as positive due to cytoplasmic staining
++ Score improved by score + 1 in three cases
Assessment in 23 casesAssessment in 23 cases
Improved H & E
NBF FAA
COST/BENEFITCOST/BENEFIT
NBF COSTLY LESS
PENETRATION TURN AROUND
IS MORE
FAA CHEAPER RAPID
PENATRATION TURN AROUND
TIME IS LESS
CONCLUSIONCONCLUSION
Though number of cases is small – a marginal benefit was observed
Results are promising and we are processing more cases
Similar study for molecular preservation in FAA Fixed tissues should be done
REFERENCESREFERENCES
1. .JOHN D. BANCROFT AND ALAN
2. STEVEN THEORY AND PRACTICE OF HISTOLOGICAL TECHNIQUES 2nd EDITION 1982
3. CARLETON H. M HISTOGICAL TECHNIQUE LONDON4. C.F.A CULLING R.T. ALLISON, W.T. BARR CELLULAR
PATHOLOGICAL TECHNIQUES 4th EDITION 1985
5. HISTOLOGICAL AND HISTOCHEMICAL METHOD BY KIERNAN 1985
6. Biotech Histochem. 1993 Mar;68(2):75-82.