The document summarizes the process for handling cytology specimens received in the laboratory. Specimens are checked for proper labeling and assigned an identification number. Conventional smears are smeared onto slides, stained, and screened. For liquid-based preparations, the sample is collected in preservative and processed using the ThinPrep 2000. This system disperses and filters cells, transferring them onto a slide which is then stained and screened. Proper handling and staining allows examination of cellular morphology.

1. When Specimens Arrive…… Check and Ensure: patient's identification on specimen tallies with that on its' request form (also known as requisition form) and the despatch form. Each case is assigned with a unique accession number and labelled clearly on the request forms as well as the specimens before processing. For Conventional Slides  CY-XXXX For Liquid Based Preparation  TP-XXXX

2. Conventional Smears Specimen is smeared onto a microscopic glass slide, spray fixed with alchohol, and sent to lab. Receive and stain the slides (using the Leica XL auto-stainer). Manual mount slides before despatching them out to the cytotechnologists on screening duties (also known as screeners) who will screen the slides for any abnormalities etc.

3. Leica XL auto-stainer Taken from: http://medequipsource.com/images/Leica%20Autostainer%20XL.jpg

4. Liquid Based Preparation Patient's gynaecological sample is collected by the clinician using a cervical sampling brush, immersed and rinsed into a vial containing PreservCytsolution. PreservCyt Solution contains methanol, which enhances cell preservation and nuclear morphology. Specimen vials will be sent to the lab. After samples are received in the lab, processing is done using the ThinPrep 2000 processor. If samples received are too mucoid or bloody, digestion must be done before running the samples using the processor.

5. ThinPrep 2000 Processor Taken from: http://websites.labx.com/rankin/pics/50949.jpg

6. Liquid Based Preparation ThinPrep 2000 Make use of mechanical, pneumatic and fluidic principles for dispersion, cell collection, and cell transfer. Accessories: Specimen Vial, ThinPrep Microscopic Glass Slide, Fixative Vial, Transcyt membrane filter, filter cap. The specimen vial, fixative vial and the microscopic glass slide (which is a special coated glass slide for preparation of ThinPrep samples) is placed into the machine.TheTranscyt membrane filter is fixed onto the filter cap before being placed into the machine.

7. Left: Brush for specimen collectionRight: Specimen Vial containing PreservCyt Solution Taken from: http://www.imvs.sa.gov.au/tissuepath/graphics/cervex_thinprep.jpg

8. Left: Conventional Smear SlideRight: ThinPrep Slide (specimen collected at the centre circle area) Taken from: http://cytologystuff.com/indexppt.htm?../powerpoint/morph1.htm

9. Liquid Based Preparation ThinPrep 2000 Process: Fluid level detection  Dispersion  Filter wetting  Cell Collection  Waste Clearing Bubble Point Cell transfer. The filter assembly (filter cap + Transcytfilter) will be lowered; specimen vial raised towards the assembly. When the membrane filter detects/makes contact with the fluid in the specimen vial  check if level is satisfactory before carrying on the process. If level of fluid is not satisfactory, it will halt the program.

10. Liquid Based Preparation ThinPrep 2000 Dispersion system activated; rotates the Transcytfilter assembly within the specimen(cell suspension)  creating shear forces in the fluid that are strong enough to separate randomly joined material and disperse mucus. Negative pressure will be applied to aid in the drawing of a small amount of fluid through the Transcytfilter to wet it.

12. Liquid Based Preparation ThinPrep 2000 The slide handler move the slide into contact with the transcytfilter  natural adhesion property of the cells and electrochemical charge of the glass slide promote the transfer of cells from the filter membrane onto the slide. Cells have a higher affinity for the glass slide than the membrane. Slight positive pressure may be introduced to aid in the process.

13. Liquid Based Preparation ThinPrep 2000 Slide will then be dropped into the fixative vial  PAP stained using auto-stainer (just like conventional smears)  manual mount and dispatched to screeners for screening. Fixative vial contains 95% ethanol, which aid in fixing the cells onto the slides, and prevent cell drop (which may occur as slides are not necessarily immediately stained.)

14. Slides placed (with label end facing down) here. Fixative vial placed here. Filter Cap Inside the processor Taken from: http://cyto.igabinet.pl/data/user_files/image/TP%20Processor%202.JPG Filter Assembly Transcyt Filter Specimen vial placed here.

15. ThinPrep Process (simplified diagram) Taken from: http://cytologystuff.com/indexppt.htm?../powerpoint/morph1.htm

16. PAP staining Papanicolaou Staining Show the differences in cellular morphology, maturity and metabolic activity. Fixation  nuclear staining  cytoplasmic staining  clearing Stains used: Haematoxylin: Nuclear stain to demonstrate the nuclear chromatin and nuclear membrane. Orange G: Cytoplasmic stain use to demonstrate keratin of squamous cells. Modified Eosin Azure: stains various cellular components.

17. PAP staining protocol

18. PAP staining protocol (continue)