In the field of microscopy, there are a variety of imaging techniques available, but determining how we proceed with imaging your samples boils down to understanding the objective-sample interface. In other words, one must decide if one will use an upright or inverted microscope. To distinguish the two microscope types, one must solely focus on the location of the objectives. The Upright microscope has its objectives opposite of the stage and will image on top of the sample. The inverted microscope has its objectives below the stage and will image from underneath the sample. Each microscope type has its own pros and cons, so to provide you with some insight as to how we choose what microscope to use, here are three questions asked to clients:
What is the thickness of your sample?
Why do we ask this? The thickness of your sample is a limiting factor in how much light can pass through. How light passes through and bounces back or refracts is key to detecting your sample and ultimately provides us with an image to analyze. At Visikol, we handle tissues from various organs and cell types for our clients. Thicker tissue may be easier to image and view through an upright microscope than an inverted one. Understanding how thick your tissue is will provide us great insight into how close and in what orientation we place it relative to the objective.
Is your sample living or fixed?
Why do we ask this? Asking if a sample is living versus fixed is more pertinent to our in vitro department, as they handle 2D and 3D cell culture, but regardless this is a question they need to ask when deciding the microscope to use for that type of imaging. For instance, a live cell culture may require them to image using an upright scope with a dipping lens, to get closer to the sample. While fixed cell monolayers may be easier to image using an inverted microscope since they are thinner than tissue and lie flat on a glass slide.
Finally, what medium is your sample in/on?
Why do we ask this? The medium that holds your sample, whether it be glass, plastic, or if your sample is in some form of aqueous solution such as culture media, can dictate the positioning of the objective as well as how close the objective needs to be to the sample.
Here at Visikol, our scientists transform a client’s experiments into images no matter the size of the tissue, living or fixed, on glass or plastic. Let us know how we can help you with your next project!