This is an article on different tips and tricks for imaging samples in a microscope. Many people using a microscope may find it challenging to acquire images or find the correct settings to use. This article will go over the different approaches you can take to increase your sample acquisition efficiency.
1. Lossless File Formats
Several lossless file formats can be used with a microscope. These file formats will produce higher-quality images and make more information about the sample visible.
GIF format: GIF is a lossless format for storing information. Compression is done on a per color channel basis, and each pixel has its data.
JPEG: JPEG is a compression method used to compress and store images and videos. It is lossy, but most people do not notice the difference between compressed images to uncompressed ones. However, there may be some artifacts from the compression process that can appear in your image.
TIFF: TIFF is another lossless compression file format that can be used for storing information. It is an image file format that was created to overcome the limits of other image formats.
2. Proper Exposure Time
The correct exposure time can be vital to an image acquisition process. Having too much light on your sample will result in overexposed images, and having less light on your sample will result in underexposed images or empty white spaces where there should be colored structures. Overexposure is when your sample receives more light than it needs, and underexposure is when there isn’t enough light hitting the sample to make it visible or a camera.
3. Focus Adjustments
As discussed previously, using the proper exposure time is essential to make an image with the correct coloration and density. In addition to taking a proper exposure, you may need to adjust the focus if you cannot find it just by finding a bright point on your sample or seeing it from far away. You can focus your lens by turning your microscope’s aperture control ring. Also, focus adjustment will help you obtain a clear focused area for image acquisition.
4. Making Sure Your Staining Is Correct
Staining can be a significant part of an image acquisition process. It will help you see structures in your sample that you previously could not see. Many stains can be used, but it is vital to ensure that the stain is compatible with the microscope and the sample you are staining. Some stains may even change or remove structures from a sample if it isn’t compatible with the stain or other chemicals in the fluid where it is being stained.
Samples can fade over time, so having a plan to preserve a sample after imaging its image is important. You should put your samples in a proper staining solution if you don’t want to stain them during imaging. Imaging should take place before you put your sample under other solutions to prevent fading of structures. Depending on the type of chemical present in your sample, different chemical solutions will be needed to prevent fading.
6. Using the Suitable Filters
With several microscopes, it is essential to know what lighting sources are compatible with your microscope scope and what filters are needed for optimal image acquisition. Different microscopes have different color modes, which can help with image acquisition, so it is important to know what color mode corresponds to your particular scope model. In addition to the color mode of your scope, you will need to adjust your camera’s settings depending on the camera attached to your microscope.
7. Use the Highest Resolution Mode for an Image Acquisition
In microscopy, it is possible to have different modes to use for imaging samples. High-resolution modes will take more time and give you better quality images, while low-resolution modes can be faster and give you lower quality images. The high-resolution images will usually look better and show more information about a sample than low-resolution images, but they take more time and effort to acquire. Sometimes the lower resolution images can be acceptable if there is a lot of information about a sample already (i.e., morphology).
8. Make Sure You Can Focus On Your Sample
Sometimes it isn’t easy to find a focal position on a sample. The microscope may be too far away from the sample, or you may not have enough contrast in your image to be able to focus. Once you focus, remember to take as many images as possible so that the specimen does not move and blur in between those images.
9. Using the Appropriate Magnification Level for Acquisition Takes Practice
Some samples will need higher magnification than others, but acquiring inspection images is very different from acquiring high-quality images of specimens. You will need to look for structures that are apparent to the naked eye or can be seen at lower magnification, then increase the magnification level on your microscope once you reach a point where you can no longer see structures on your sample, decrease the magnification level to obtain high-quality images of your sample.
10. Be Patient With Your Microscope
Every microscope is different and has its quirks, so it may take a while to acquire a high-quality image. It can also take a while to be able to acquire a good image of a sample. Patience is one of the essential skills that you will need as an imaging scientist.
The list above is by no means exhaustive, but it should give you a basic overview of the most common mistakes that imaging scientists make while acquiring images. These mistakes can prevent you from achieving a high-quality image of your sample. Remember that even if you did acquire a good image, there may be many problems with the image due to other factors such as exposure time and focus adjustments. Finding these problems will allow you to fix the problems in your images to make them better. When choosing an expert to process your images, it is essential to know what kind of equipment they have and what limitations they have to get the best results possible.