Preparation Of A Tissue Imprint Using A Cytological Device

The CerviSoft cell preparation technique has a substantial advantage over touch preparation in that cell density is better controlled. The formation of large clusters, which are not recognized by a laser scanning cytometer (LSC), is significantly reduced (Bollmann et al., 2002). This protocol requires a CerviSoft Foam device (Puritan Medical Products); for other needed materials, see Support Protocol 1.

1. Prepare microscope slides and a solid tumor sample as described (see Support Protocol 1, steps 1 and 2).

2. Cut a fresh tumor surface as large as possible (10 to 20 mm in diameter).

3. Roll the sponge part of a CerviSoft device over the freshly cut surface in one direction.

For details on using and maintaining the CerviSoft device, see manufacturer's instructions and Bollmann et al. (2002).

4. Transfer the attached cells to slides by rolling the device in the opposite direction.

This results in an ~15 x 30-mm field that is evenly covered with cells. This area is large enough for at least two measurements: one for immunofluorescence analysis and one for the isotype control.

With one pass of the device over the tumor sample, two or three slides can be prepared before the device is discarded.

5. Dry slides and use a grease pencil to delineate individual fields.

For optimal results, slides should be stained within 2 hr of their preparation. Slides can be stored several days in plastic containers at room temperature or up to several months at -20° C.

Analysis of Tissue Imprints by Scanning Laser Cytometry

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