To prepare the biopsy tissue for wax impregnation and for further processing of paraffin block formation.
2.0 SCOPE
This sop shall be applicable to all types of tissue after initial fixation, grossing, and decalcification where applicable.
3.0 RESPONSIBILITY
3.1 User Department Person.
3.2 User Department Head.
3.3 10% formal saline preparation. Take the volume to 900 ml with distilled water and add 100 ml of 40% formaldehyde solution. Mix the solution wall store at room temperature
3.4 Quality Assurance Person
3.4.1 To execute review processes for SOP.
3.4.2 To maintain the original copy of the SOP.
3.4.3 To issue a control copy of the SOP.
3.4.4 To maintain the issuance record of SOP.
3.4.5 To retrieve the control copy of the SOP & maintain its record.
3.5 Quality Manager
4.5.1 To approve new or revised SOP.
4.5.2 To ensure the implementation of the defined system.
4.5.3 To assign work to QA executive & department.
4.0 DISTRIBUTION
4.1 The Quality Assurance department is responsible to keep SOP ‘Master Copy’ approved through Quality Assurance Head.
4.2 The copy of ‘Control Copy’ of SOP is being distributed in user departments and placed near related Equipment/Instruments as Display Copy.
5.0 DEFINITION(S)
5.1 Definition(s)
N/A
6.0 ABBREVIATION(S)
6.1 Abbreviation(s)
6.1.1 SOP: Standard Operating Procedure
6.1.2 No. : Number
6.1.3 Dept. : Department
6.1.4 QA: Quality Assurance Person or Executive
6.1.5 Next Rev: Next Revision
6.1.6 Rev. : Revision/Review
7.0 PROCEDURE
7.1 Analytical steps
7.1.1 The desired program of the processing is selected
7.1.2 Start button of the instrument is switched on.
7.2 Instrument details: -----------------------------
7.3 Type of sample
7.3.1 Tissue Sample
7.4 Requirements
7.4.1 Fixed Tissue
7.5 Reagents
7.5.1 10%-formalin
7.5.2 60%-alcohol
7.5.3 70%-alcohol
7.5.4 80%-alcohol
7.5.5 90%-alcohol
7.5.6 Absolute alcohol
7.5.7 Xylene
7.5.8 Paraffin wax
7.6 Reagent preparation & storage
7.6.1 Formal-sodium acetate (stock)
Formaldehyde, 30%-40% - 10.0 ml
Sodium acetate - 2.00 gm
Tap water - 90.0 ml
7.6.2 Formal- glycerol working solution
Formalin- sodium acetate, stock- 90.0 ml
Glycerin (glycerol) - 10.0 ml
7.7 Test procedure for tissue processor
Overnight schedule (routine specimen processing (Total processing time 14-16 hours: minimum fixation time 12 hours)
7.7.1 10%-formalin ---1:30 hour-1st bucket
7.7.2 10%-formalin ---1:30 hour-2nd bucket
7.7.3 10%-formalin ---1 hour-3rd – bucket
7.7.4 60%-alcohol ---1 hour-4th – bucket
7.7.5 70%-alcohol ---1 hour-5th - bucket
7.7.6 80%-alcohol ---1 hour-6th - bucket
7.7.7 90%-alcohol ---1 hour-7th - bucket
7.7.8 absolute alcohol---1 hour-8th - bucket
7.7.9 xylene ---1 hour-9th - bucket
7.7.10 xylene ---1 hour-10th - bucket
7.7.11 paraffin wax ---1:30 hour-11th-bucket
7.7.12 paraffin wax ---1:30 hour-12th- bucket
7.7.13 Agitation of a basket containing tissue cassettes is helpful in ensuring proper penetration of reagents in the tissue and results in adequately processed tissue.
7.7.14 There are no advantages to taking shortcuts in processing. The results can be poorly and incompletely dehydration, cleared and infiltrated specimens, which will cause further problems when sectioning, staining, and examining the tissue.
7.7.15 The reagent solutions shall be changed depending upon the quality and the quantity of the samples processed. Symbolically with a load of 300-500 blocks per day, the solution need to be changed as follows: absolute alcohol (1st container)-alternate day, xylene –every 3rd day, and wax weekly .however, formalin needs to be changed every day.
7.7.16 The log shall be created and maintained by the technician for the days of changing the processing solutions.
7.8 Method for reprocessing tissue procedure
7.8.1 Place block or specimen in molten paraffin for 1 hour
7.8.2 Xylene, 3 changes 1 hour each
7.8.3 Absolute alcohol 2 changes, 1 hour each
7.8.4 95% alcohol, 2 changes, 1 hour each.
7.8.5 Running tap water for 30 minutes.
7.8.6 Place in the formal-glycerol solution until tissue becomes soft and pliable with gentle pressure.
7.8.7 Reprocess on tissue processor in the usual manner.
7.9
Maintenance of tissue
processor
|
TASK |
FREQUENCY |
|
Cleaning Of Display And Keyboard |
Daily |
|
Cleaning Of Basket Lid |
Daily |
|
Cleaning Of All Chemical Containers |
weekly |
|
Cleaning Of Residual Paraffin From The Surface Of The Station Holder And
Lid Holder. |
Daily |
|
Cleaning Of External And Internal surfaces |
weekly |
8.1 All specimens should be handled with gloves.
8.2 All instruments should be handled with care to prevent injury.
9.0 REFERENCES
9.1 References Refer to the manufacturer’s manual for additional information.
END OF THE DOCUMENT
You may like to read these links:
1. List of All SOPs and Documents for the Microbiology Laboratory
2. List of All SOPs and Documents for In-vitro Laboratory
3. List of All SOPs and Documents for the In-vivo Laboratory.
4. List of All SOPs and Documents for Clinical Research
5. List of Instruments and Equipment Used In Laboratory Animal Facility
6. List of All SOPs and Documents for the Animal House Facility
7. List of SOPs and Documents for Histopathology Laboratory
8. Checklist for Toxicology (OECD-407) study preparation for Histopathology
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