Preparing Tissue Cultures for Confocal Microscopy

Preparing Tissue Cultures for Confocal Microscopy

Researchers have been fascinated with the microscopic world for millennia. Centuries ago, scientists like Robert Hooke and Antoine van Leeuwenhoek developed the first light microscopes. These devices evolved into intricate technologies that, through magnification, give researchers access to the billions of cells comprising our bodies and the living world. Among these contraptions is the confocal microscope.

Produced in the 1950s, the confocal microscope has since produced crisp, colorful images that depict living cells’ morphologies. The high resolution that confocal microscopes afford has, in turn advanced cell biology, biochemistry, and molecular genetics research. Insights from these fields have helped researchers develop novel therapeutics that improve health outcomes and quality of life.

But to best harness confocal microscopy, researchers must consider the many confocal microscopes available and develop workflows that generate clear images and reproducible data. At its core, confocal microscopy involves multiple steps, starting with tissue cultures and ending with image analysis. 

In this article, we will cover confocal microscopy and the process of preparing cell cultures for visualization. Then, we will discuss what researchers should consider when conducting a confocal microscopy study and show how our leasing program helps you acquire the equipment you need to visualize all kinds of cell lines and cell types.

What Is Confocal Microscopy?

Confocal microscopy is a fundamental visualization technique that observes tissues and samples in a three-dimensional (3D) space, facilitating 3D imaging. Marvin Minsky invented the device to address the primary shortcoming of widefield microscopy: its pinhole introduces out-of-focus light while capturing images, reducing the image’s resolution

Confocal microscopy is a kind of optical sectioning microscopy, which allows microscopes to capture high-resolution images from tissue without having to cut the tissue into slices. Two kinds of confocal microscopes exist for confocal imaging:

Like a widefield microscope, confocal microscopy can also be conducted with three kinds of imaging views.

Sample preparation of cell cultures for confocal microscopy

Before researchers can view cell cultures for confocal microscopy, living cells must first be prepared for visualization. This process features a series of steps and diverse reagents that yield quantitative image data. Here are the steps in order:

Along the pipeline, researchers must make sure to use strong microscopy techniques in their experiments. Doing so will help researchers retain cellular morphology and activity as they visualize their samples.

Applications of confocal microscopy

Confocal microscopes are ideal for studying cellular processes, from the molecular level to the level of the whole organism. With tissue cultures, researchers can characterize different kinds of tissues and cells from diverse specimens:

Researchers have also used confocal fluorescence microscopy to develop 3D tissue culture models. These structures — spheroids and organoids — have the potential to complement animal models in biomedical research by recapitulating in vivo tissue-like structures and functions. Here are just some of the ways researchers have captured human physiology with microscopy:

Considerations for confocal microscopy

As researchers begin considering a confocal microscope for visualizing cells, they must consider several visual phenomena. Each of these occurrences affect the final images and the quantitative data generated from them.

Excedr leases many kinds of confocal microscopes

Excedr has a long history of leasing confocal microscopes from many different brands. And because we do not carry an inventory, you can select the exact manufacturer and model you want to lease, specific to your experimental needs. Here are just some of the confocal microscopes we have leased in the past:

  • Zeiss: The LSM 980 and LSM 900 provide 3D and 4D imaging for fluorescently stained tissue cultures and samples. Both provide high signal-noise ratios and improved multi-color resolution to maximize image quality and produce valuable spatial information through LSM Plus. Both also provide multichannel (the LSM 900 — 3 and the LSM 980 — 36) capabilities to observe a wide array of different fluorescent stains concurrently. Lastly, these Zeiss microscopes take up a small lab footprint, minimizing the lab space and time required for user training.
  • Keyence: The company produces two kinds of CLSMs: the VK-X series and the BZ-X series. VK-X harnesses surface metrology to resolve fluorescence structures 12 times faster and 16 times larger than conventional microscopes. The BZ-X is a fully motorized confocal microscope system that supports fluorescence, brightfield, and phase-contrast imaging as well.
  • Olympus: Olympus boasts multiple types of CLSMs but the FV3000 is specifically designed for life sciences research. The microscope achieves 3 times more light transmission than traditional spectral detection technologies with the TruSpectral technology. The microscope also comes equipped with lambda scanning mode to tease apart overlapping fluorescent signals. Automation with the FV3000 is also possible thanks to a Macro-to-Micro module in Olympus’s software.

Lease your confocal microscope with Excedr

Confocal microscopes provide researchers with high-resolution digital images of microscopic organisms. Such images provide an unprecedented view into the cells that help our bodies function. 

Understanding cell biology with a confocal microscope also helps researchers develop pharmaceutical and biologics to advance biomedical research. While researchers must consider many factors and refine sample preparation pipelines, doing so will help end users make the best use of the confocal microscopes available.

Excedr’s leasing program can help advance cell biology research for your lab. Speak with our team today to learn how we can help you acquire confocal microscopes that match your needs.