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Microscopy School Lesson 7 - Sample Preparation for Immunohistochemistry

Microscopic images are amazing; they allow us to see deep inside cells and tissues uncovering a world that otherwise would be hidden. Nevertheless, the outstanding image that represents the results clearly and with high quality relies on two key factors: 1) acquisition with the appropriate equipment and, 2) excellence in sample preparation. Neither of those factors can be disregarded because, if so, the final result will be deeply compromised.

Regardless of the equipment, sample preparation is a critical factor that is often disregarded by less experienced microscopists. In fact, irrespective of the quality of the imaging equipment, poor quality samples will always deliver a poor-quality image. Therefore, sample preparation is crucial for the overall imaging quality.

There are several steps for Immunohistochemistry sample preparation. It begins with adequate sample collection and fixation. Subsequently, the samples are stained with antibodies. Antibody staining is crucial and dependent on several factors that must be determined experimentally.

In recent years together with the increasing microscopy power to image large samples, several post-treatment clearing techniques have emerged, allowing in toto imaging of fluorescently labelled samples. Currently, all these new technological advances enable the imaging of biological samples with an unprecedented morphological contextualization.

In this webinar, Dr Marco Campinho (CBMR - Universidade do Algarve) discusses every aspect of sample preparation for Immunohistochemistry.

Learning Objectives:

  • List the critical steps in sample preparation for Immunohistochemistry. 
  • Recognize the caveats in sample preparation.
  • Design and plan an immunohistochemistry experiment.

Questions Answered: 

  • What are the critical steps in Immunohistochemistry sample preparation? 
  • How to optimize antibody staining? 
  • What is clearing?

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