Vet360 Vet360 Vol 05 Issue 01 | Page 27

CPD ACCREDITED ARTICLE 3. Scrapings Scrapings could be useful for obtaining cells from firm surfaces, such as firm cutaneous lesions. Cells obtained by means of a scalpel blade are gently smeared onto a clean glass slide. This often gives many broken cells, but areas of intact cells can usually be found. 4. Swabs Swabs can be used to obtain cells from mucosal surfaces such as the rectum or vagina, or in areas where the other sampling methods are not practical, such as fistulous tracts. The swab must be gently rolled along the surface of a clean glass slide. Preparation of slides Once a sample has been collected, it needs to be transferred to a clean glass slide and spread out - the aim is to obtain a monolayer of cells. This needs to be done quickly before the material dries out or clots. There are various techniques, depending on the volume and consistency of collected material. 1. Solid tissue aspirates • Slide over slide (“squash”) preparation This is generally the best method for preparing slides, if done properly but does need some practice. The material collected is placed in the middle of a clean glass slide (the smear slide). A second slide (the spreader slide) is placed over the sample, perpendicular or parallel to the smear slide. The weight of the spreader slide will cause the sample to spread. Once this has happened, the spreader is gently drawn across the smear slide, without any downward pressure, as this will rupture cells. (Fig 3) The weight of the second slide is enough to cause the cells to spread. Non-fragile tissues (e.g. from carcinomas) or any thick viscous material (such as mucous strands from trans-tracheal aspirates) can be spread by this method. With some experience, and a gentle touch, this method can also be used for more fragile tissues like lymph node or spleen. Fig 3: Slide over slide (“squash”) preparation Fig 4: Blood smear technique • Blood smear technique This is done in a similar manner to making a blood smear. The material is placed near the end of a smear slide. The spreader slide is placed in front of the material at a 45-degree angle, backed up until the material is touched and then moved forward. (Fig 4) The feathered edge should not be right on the edge of the slide, as the stage retaining clamps on the microscope will interfere with the oil immersion lens and the sample then cannot be viewed. Malignant cells are often found in the feathered edge. This technique works well for tissues with a fluid component, like internal organs and lymph nodes. • “Starfish” preparation Here the tip of a needle is used to drag the material in several different directions. It is a gentle technique that does not cause much rupture of the cells, but it does leave a thick layer of tissue fluid around the cells that may stop them from spreading to an acceptable shape and size.(Fig 5) Usually some areas that can be interpreted are found. This is not a commonly used or preferred technique (rather try a squash prep), but can be used when grainy or viscous material is aspirated. Fig 5: “Starfish” preparation Issue 01 | MARCH 2018 | 27