How can you reduce artifacts in microscopy images?

1 Sample preparation

The first step to avoid artifacts is to prepare the samples properly for the microscopy technique you are using. Depending on the method, you may need to cut, polish, coat, embed, or stain the samples to make them suitable for observation. However, these processes can also introduce damage, contamination, or contrast variations that can affect the image quality. To reduce these artifacts, you should follow the recommended protocols for each technique, use appropriate tools and materials, and minimize the exposure of the samples to heat, air, or chemicals.

2 Alignment and calibration

The second step to avoid artifacts is to align and calibrate the microscope and its components before taking the images. Misalignment or miscalibration can cause blurring, distortion, noise, or aberrations in the images. For example, if the objective lens is not centered, focused, or clean, it can produce uneven illumination, chromatic aberration, or dust spots. To reduce these artifacts, you should check and adjust the alignment and calibration of the microscope regularly, following the manufacturer's instructions and using standard reference samples.

3 Image acquisition

The third step to avoid artifacts is to optimize the image acquisition settings and conditions for the microscopy technique you are using. Different techniques have different parameters that affect the image quality, such as magnification, resolution, exposure, contrast, wavelength, or voltage. If these parameters are not set properly, they can cause overexposure, underexposure, saturation, aliasing, or moiré patterns in the images. To reduce these artifacts, you should choose the optimal settings and conditions for your samples and objectives, and avoid changing them during the image acquisition.

4 Image processing

The fourth step to avoid artifacts is to apply appropriate image processing techniques to enhance the image quality and extract useful information. Image processing techniques can include filtering, sharpening, denoising, deconvolution, segmentation, or quantification. However, these techniques can also introduce artifacts if they are not applied correctly or excessively. For example, if the filter size is too large or too small, it can remove important details or create false edges. To reduce these artifacts, you should use image processing techniques that are suitable for your microscopy technique and data, and avoid overprocessing or manipulating the images.

5 Image interpretation

The fifth step to avoid artifacts is to interpret the images correctly and critically, taking into account the limitations and assumptions of the microscopy technique and the image processing techniques. Sometimes, artifacts can be mistaken for real features or properties of the samples, or vice versa. For example, if the contrast is too high or too low, it can create false positives or false negatives. To reduce these artifacts, you should compare the images with other sources of information, such as literature, simulations, or complementary techniques, and validate your interpretations with experiments or tests.

6 Image presentation

The sixth step to avoid artifacts is to present the images clearly and honestly, following the ethical and professional standards of the materials science community. When you present the images, you should include the relevant information about the microscopy technique, the sample preparation, the image acquisition, and the image processing. You should also indicate any modifications, enhancements, or annotations that you have made to the images, and explain their purpose and rationale. To reduce the artifacts, you should present the images as they are, without exaggerating, omitting, or falsifying any data.

7 Here’s what else to consider

This is a space to share examples, stories, or insights that don’t fit into any of the previous sections. What else would you like to add?