Changing your IHC/ISH stainer – what to consider

The mission of the Princess Maxima Centre for Children’s Oncology is to cure every child with cancer, with an optimal quality of life. Close collaboration between care and research will lead to better patient management for children with cancer.

This means that all patients (with permission, of course) participate in research and all results of the research done is readily available for all future patients. This is essential for better patient management of children with fewer side effects and late effects.

Ella de Boed a team leader in Pathology shared the core values off the Princess Maxima Centre for Children’s Oncology are: Ground-breaking and Passionate.

The Laboratory for Children Oncology (LKO) consists of five different departments (Biobank, Morphology, Flowcytometry, Molecular and Pathology) which work closely together to provide the best diagnosis for our patients.

The Pathology lab of the LKO processes the entire spectrum of the children’s diseases, as well as performing research activities for both its own and external patients. The main goal of the Research department is to improve patient care. Quality of education, training and refresher courses are paramount to this.

The current challenges lie in setting up new antibodies, especially biomarkers to determine which immunotherapy the patient should get. Also implementing non-standard antibodies, especially for brain tumours, and double staining with non-standard antibodies provide some challenges. In the future we also want to implement IHC-staining on organoids.

The goal is to provide the clinic with a highly confident diagnosis as soon as possible, to start treatment of the child patient as soon as possible.

Our goal is to minimize the number of different protocols on the Leica BOND-III, as this might cause some delay. Both Clinic and Research work close together to formulate better therapies and diagnoses for the children, resulting in a higher chance of survival.

The Princess Maxima Centre for Children’s Oncology is a new hospital, and the IHC-staining had been performed by the University Hospital next door. As time progressed the need to have IHC-staining capabilities in-house grew, resulting in the decision to start the procurement procedure for an IHC-stainer.

The decision to buy the Leica BOND-III Immunostainer was made after a careful selection process, where multiple criteria were scored and compared between the different vendors.

During the selection process multiple slides were also stained on the different staining platforms, and the quality of the staining was compared.

The scoring of the criteria combined with the staining quality, combined with the quotes, resulted in our choice to purchase the Leica BOND-III stainer.

The choice to purchase the BOND was made based on several items. The goal was to perform as many slides as possible with a small footprint. The instrument needed to be robust, user friendly and preferably the waste needed to be separated. Due to the waste separation the amount of hazardous waste needed to be as low as possible.

A bi-directional connection between the staining platform and our LIS-system was also a key requirement. Also, as we are treating children, we would like to be able to have a short turn-around-time and be able to dehydrate plus coverslip the slides as soon as they are done.

The Princess Maxima Centre for Children’s Oncology is a new hospital, and the IHC-staining had been performed by the University Hospital next door. Whilst the new IHC-lab was set-up, and the lab techs were hired, the procurement procedure for the IHC-stainers was already ongoing.

There was a project plan/action plan set up to align all steps in this process, and during this time we established the IHC-department in the Princess Maxima Centre for Children’s Oncology from scratch.

Here are the key areas covered by the project plan

1. Check the validation control material required is available and ready to cut for each antibody on the list to validate
2. Check there is enough material to perform the validation given the number of slides for each antibody and controls required
3. Establish the protocol for the intra run on the IHC//ISH stainer
4. Establish the staining acceptance criteria
5. Once the optimum protocol for each antibody is determined that protocol for the specific antibody is set up on the each IHC/ISH stainer for future use

Lab workflows
The workflow is now optimized. Before we started our own IHC-department, the slide was stained at the University Hospital next door. Now that we perform IHC-staining in-house, the Turn-Around-Times have improved enormously. We also encounter fewer errors due to the bi-directional LIS-connection.

Patients and the clinic?
We can provide our patients with a diagnosis much faster, which means that the therapy for the patient can start quicker. Today we provide a 24 hrs IHC service and previously it took 1 to 3 days.

For In Vitro Diagnostic Use
¹KIKA Children Cancer Free Foundation. Available at www.kika.nl/children-cancerfree-foundation. Accessed June 26, 2023