SIP slides on CYTO - gynae

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SIP slides on CYTO - gynae

  1. 1. When Specimens Arrive……<br />Check and Ensure:<br />patient&apos;s identification on specimen tallies with that on its&apos; request form (also known as requisition form) and the despatch form. <br />Each case is assigned with a unique accession number and labelled clearly on the request forms as well as the specimens before processing.<br />For Conventional Slides  CY-XXXX<br />For Liquid Based Preparation  TP-XXXX<br />
  2. 2. Conventional Smears<br />Specimen is smeared onto a microscopic glass slide, spray fixed with alchohol, and sent to lab. <br />Receive and stain the slides (using the Leica XL auto-stainer).<br />Manual mount slides before despatching them out to the cytotechnologists on screening duties (also known as screeners)<br />who will screen the slides for any abnormalities etc. <br />
  3. 3. Leica XL auto-stainer<br />Taken from:<br />http://medequipsource.com/images/Leica%20Autostainer%20XL.jpg<br />
  4. 4. Liquid Based Preparation<br />Patient&apos;s gynaecological sample is collected by the clinician using a cervical sampling brush, immersed and rinsed into a vial containing PreservCytsolution.<br />PreservCyt Solution contains methanol, which enhances cell preservation and nuclear morphology.<br />Specimen vials will be sent to the lab. After samples are received in the lab, processing is done using the ThinPrep 2000 processor. <br />If samples received are too mucoid or bloody, digestion must be done before running the samples using the processor.<br />
  5. 5. ThinPrep 2000 Processor<br />Taken from:<br />http://websites.labx.com/rankin/pics/50949.jpg<br />
  6. 6. Liquid Based Preparation ThinPrep 2000<br />Make use of mechanical, pneumatic and fluidic principles for dispersion, cell collection, and cell transfer.<br />Accessories:<br />Specimen Vial, ThinPrep Microscopic Glass Slide, Fixative Vial, Transcyt membrane filter, filter cap.<br />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.<br />
  7. 7. Left: Brush for specimen collectionRight: Specimen Vial containing PreservCyt Solution<br />Taken from:<br />http://www.imvs.sa.gov.au/tissuepath/graphics/cervex_thinprep.jpg<br />
  8. 8. Left: Conventional Smear SlideRight: ThinPrep Slide (specimen collected at the centre circle area)<br />Taken from:<br />http://cytologystuff.com/indexppt.htm?../powerpoint/morph1.htm<br />
  9. 9. Liquid Based Preparation ThinPrep 2000<br />Process: Fluid level detection  Dispersion  Filter wetting  Cell Collection  Waste Clearing Bubble Point Cell transfer.<br />The filter assembly (filter cap + Transcytfilter) will be lowered; specimen vial raised towards the assembly. <br />When the membrane filter detects/makes contact with the fluid in the specimen vial  check if level is satisfactory before carrying on the process. <br />If level of fluid is not satisfactory, it will halt the program.<br />
  10. 10. Liquid Based Preparation ThinPrep 2000<br />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.<br />Negative pressure will be applied to aid in the drawing of a small amount of fluid through the Transcytfilter to wet it.<br />
  11. 11. Liquid Based Preparation ThinPrep 2000<br />System will gently blow out the liquid in the filter to clear any cellular material from the filter menbrane surface<br /><ul><li>membrane is actually a biologically neutral, flat, smooth and porous surface which is capable of collecting cellular material.</li></ul>Pneumatic system will apply positive and negative pressure to the filter in a series of pulses draw the specimen through the filter membrane and collect the suspended cellular material onto the membrane surface. <br /><ul><li>As material in the specimen vial collects onto the filter mebrane, the pores of the membrane gets blocked, thereby triggering the processor to stop aspirating.</li></li></ul><li>Liquid Based Preparation ThinPrep 2000<br />Collection ends  transcytfilter is withdrawn from the specimen vial  the filtrate is aspirated into the waste bottle as the filter is inverted; collected cells will remain of the filter due to negative pressure.<br />Excess fluid will be removed from the filter membrane before transferring the cells onto the glass slides  to enhance cell adhesion to the slides. <br />
  12. 12. Liquid Based Preparation ThinPrep 2000<br />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.<br />Cells have a higher affinity for the glass slide than the membrane.<br />Slight positive pressure may be introduced to aid in the process.<br />
  13. 13. Liquid Based Preparation ThinPrep 2000<br /> 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.<br />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.)<br />
  14. 14. Slides placed (with label end facing down) here.<br />Fixative vial placed here.<br />Filter Cap<br />Inside the processor<br />Taken from:<br />http://cyto.igabinet.pl/data/user_files/image/TP%20Processor%202.JPG<br />Filter Assembly<br />Transcyt Filter<br />Specimen vial placed here.<br />
  15. 15. ThinPrep Process (simplified diagram)<br />Taken from:<br />http://cytologystuff.com/indexppt.htm?../powerpoint/morph1.htm<br />
  16. 16. PAP staining<br />Papanicolaou Staining<br />Show the differences in cellular morphology, maturity and metabolic activity.<br />Fixation  nuclear staining  cytoplasmic staining  clearing<br />Stains used:<br />Haematoxylin: Nuclear stain to demonstrate the nuclear chromatin and nuclear membrane.<br />Orange G: Cytoplasmic stain use to demonstrate keratin of squamous cells.<br />Modified Eosin Azure: stains various cellular components.<br />
  17. 17. PAP staining protocol<br />
  18. 18. PAP staining protocol (continue)<br />
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