BLT Stages

In Situ hybridization is a very important technique in cancer research. The aim of this project is to perform an optimization and validation of the mouse double minute 2 (MDM2) in situ hybridization on the Ventana Benchmark GX. This test is executed to differentiate between a lipoma and a well differentiated liposarcoma (WDLPS)/atypical lipomatous tumor or between a malignant tumor and a dedifferentiated liposarcoma (DDLPS).

The MDM2 in situ hybridization is a new emerging test, that is why this needs to be compared with the fluorescence in situ hybridization (FISH). The optimization of the test is being done with the standard protocol, testing 12 slides where there are 6 positives and 6 negatives. Further optimization is needed when the acquired signals are not strong enough. Further optimization was not needed in this test. The test is validated using an inter and intra-run with 6 samples. In the inter-run 1 slide per sample is tested in a different run with the standard protocol to calculate reproducibility of the test. With the inter-run a comparison is made between different lot numbers. To calculate the repeatability, 3 slides are tested from the same 6 samples in the same run.

The interpretation of the results is based on the MDM2/CHR12 ratio, for which a score sheet has been created. The reproducibility and repeatability are calculated by determining the coefficient of variation between the averages of MDM2/CHR2 ratios. The obtained coefficients of variation are low, so that there is good reproducibility and repeatability.

The conclusion of this optimization and validation is that this test has good reproducibility and repeatability. As a result, this test will be carried out in practice at the Jan Yperman Hospital.

Cervical cancer is the second most common cancer in women in the world, while it is the leading cancer in women in the developing countries. Unlike most other malignancies, cervical cancer is readily preventable because it has a long preinvasive phase. Cervical cytology has as goal to detect women with epithelial abnormalities. This study examines how the cervical cytology is performed.

This test is used at the Jan Yperman Hospital. It’s laboratory anatomo-pathology has recently bought a new staining device (Compass Stainer of Hologic) to perform the Papanicolaou staining. This device has to be validated before it can be used.

In the validation process, it is tested whether the coloration is the same as that of the previous staining device (TST44 of Medite). It is checked whether the staining is equal to a sample that is processed several times in one run (repeatability). When a sample is processed several times is different runs, the result has also to be the same (reproducibility).

The Compass Stainer meets the requirements and is therefore validated. The staining is equal as the staining of the previous staining device. It is also repeatable and reproducible. The Compass Stainer is therefore applicable today at the Jan Yperman Hospital.

The laboratory of pathology-located in the Jan Yperman hospital, is the central key in the project called ‘Pegasus’. This is a project amongst different laboratories of pathology in Flanders. The intention of this project is to speed up the process of giving second opinions-about tumoral tissues. The key instrument is a digital scanner. The scanner is able to make whole slide images. Before the project starts, the scanner has to be validated.

First of all, a SOP is written so everyone uses the scanner the same way. For the validation of the scanner, the guidelines of the College of American Pathologist have been used. Twenty slides,where a diagnose with microscope is already made a couple of weeks before, are scanned. Dr. Kristof Cokelaere makes a diagnosis on a monitor.The diagnosis on a monitor and on a microscope has to be the same. A couple of things need attention. The quality of the image, the completeness of the image, readability of the barcode, etc.

The quality of the image and the readability of the barcode are accurate. There are problems with the completeness of the image. If the original tissue isn’t at the middle of the slide, the scanner can’t scan it right. The image is out of focus. The most important thing is the quality of the original slide. Different thickness of the tissue, air bubbles, spots, etc. can create a bad whole slide image.

For this project, the scanner can be used-if a first diagnosis had already been made. The scanner can’t be used for daily research.