Standard Operating Procedure (SOP) for Processing of Cytology Fluids
1.0 Purpose:
To establish a standardized procedure for the processing of cytology fluids within the Cytopathology section.
2.0 Scope:
This procedure applies to all types of fluids, fine needle aspirations (FNA), and smears.
3.0 Introduction:
The objective of fluid cytology staining is for routine cytology analysis.
4.0 Principle:
Prior to staining, the smear must undergo fixation.
5.0 Analytical Measuring Range and Limit of Detection:
Not applicable.
6.0 Responsibility:
Technical staff and pathologists are responsible for adherence to this procedure.
7.0 Abbreviations:
None.
8.0 Requirements:
Kit Reagents:
Alcohol with varying gradations (70%, 90%, 95% absolute).
Centrifuge
Slides
Cover slip
Diamond pencil
Cyto centrifuge
Instrumentation and Software:
Centrifuge
Slides
Cover Slip
Diamond pencil
Cyto Centrifuge
Centrifuge Tube
Disposables:
Gloves
Specimen Collection and Handling:
a. Specimen Collection: Slides must be fixed immediately
after smear preparation at room temperature.
b. Specimen Transport: At room temperature.
c. Specimen Storage: At room temperature.
d. Specimen and Control Preparation: Not applicable.
9.0 Precautions:
All specimens should be handled with gloves.
Instruments should be handled carefully to prevent injury.
10.0 Limitations & Interferences:
Not applicable.
11.0 Documentation:
All documents are maintained in:
Cytology entry register
Cytology stain adequacy form
Cytology reagent change form
12.0 Preparation of Slide for Cytology:
Cytocentrifuge:
Take one drop of fluid in each of the four cytospin tubes
for 10 minutes at 1000 RPM.
Fluid:
Take fluid in a 15 ml tube and centrifuge at 2500 to 3000
RPM for 10 minutes.
Make four smears on clean slides:
a) Giemsa
b) PAP smear
c) H & E stain
d) One extra smear.
Fix slides in 95% ethanol or methanol.
13.0 Instructions:
13.1 Pre-Analytical Steps:
All specimens are entered into the Cytology Entry Technical
register and assigned a department ID.
The same numbers are noted during all processing steps for
specimen integrity.
13.2 Analytical Steps (Method for PAP Stain):
Wash smear with running tap water for 2 to 5 minutes.
Stain with Hematoxylin for 3 minutes (Conventional) or as
per LBC protocol.
Wash with running tap water for 2 to 5 minutes.
Differentiate with 0.5% aqueous Acid Alcohol for 5 to 10
seconds and wash.
Blue in 0.1% aqueous ammonia solution for 5 to 10 seconds
and wash.
Stain with OG6 for 4 minutes (Conventional) or as per LBC
protocol.
Clear with xylene for 2 to 3 minutes.
Mount with DPX and label the slides.
Adjust staining times based on preference.
Submit slides to the pathologist along with the requisition
form.
13.3 Quality Control and Assurance:
Evaluate a suitable slide of each type for staining adequacy
before each batch.
13.4 Calibration and Calibration Verification:
Not applicable.
13.5 Storage of Samples:
Cytology samples are retained for 10 years at room
temperature.
Fluids are stored at 2-8 degrees Celsius in a refrigerator
for 7 days.
14.0 Reference Range:
Not applicable.
15.0 Critical Value:
Report critical/significant findings according to SOP reporting standards.
16.0 Reporting of Results and Interpretation:
Results are reported following the established Bethesda Protocol in approved report formats.
17.0 Contingency Plan:
Suitably trained staff are available as backup in case of reagent shortages, manpower shortages, or instrument breakdowns.
18.0 References:
Solomon D. Nayar R. Eds. The Bethesda system for Reporting
cytology. Definitions criteria and Explanatory Notes, 2nd Ed. 2004.
Ramnik Sood, Medical Laboratory Technology. Methods and
Interpretations, 6th ed., 2009.
END OF THE DOCUMENT
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