In our previous blog post we discussed resonant and polygonal mirror scanning systems. These scanning systems are designed for high-speed imaging, and can achieve short pixel dwell times compared to conventional galvanometer mirrors. While short pixel dwell times...
In our previous post, we discussed galvo scanning systems used in laser scanning confocal and multiphoton microscopes. Galvo scanning is standard, yet for some applications, such as live-cell and large-format image acquisitions, systems that offer increased...
Taking the time to understand the inner workings of an imaging system can help you design more robust experiments and acquire higher-quality data. It can also help you trouble-shoot when a system is not performing as expected. Over the next few posts, we will...
In the last post, we reviewed how scattering and absorption attenuate the intensity of the laser used to excite fluorescence and reduce the amount of emission signal that can make it to the detector(s). Tissue is very complex, and fluorescence from different dyes, as...
When carrying out fluorescence imaging in thicker samples, we often hear about how “scattering” affects image quality by limiting the depth of imaging within the sample. Scattering occurs when light traveling through a sample encounters a change in refractive index, ...
Often we want to visualize multiple structures within the same preparation, and if the colors are not registered, this can lead to erroneous conclusions. For example, if you are counting nuclei, the color correction does not likely matter, but when examining whether a...
Over the next few posts, we will dig deeper into microscope objectives and explore how they can affect the quality of your imaging. Let’s start the series off with magnification and resolution, which can be a point of confusion when planning experiments. Often,...
The meaning and relevance of bit depth is a common point of confusion for life scientists who use microscopes. Typically, when we mention bit depth in microscopy, we are referring to gradations in the intensity readings from a detector. Let’s use a photomultiplier...
The main advantage of two-photon microscopy is the ability to image deeper into scattering samples, such as tissue and live specimens. One way that two-photon objectives assist deep imaging is by providing extended working distances compared to lenses for other...
In our last post, Craig and I introduced the basics of two-photon microscopy. This month we will review some considerations for choosing the best objective for your multiphoton imaging. If you are new to objectives and their properties, we recommend that you consult...