2014 Most Popular Microscopy Thesis

What's more amazing than the image microscopists get from the cellular world? In addition to the United States, such photographs also reveal new insights into the "way of how cells and biomolecules operate and interact." What research progresses in microscopy technology in 2014 made us greatly admired?

By imagination alone, we cannot see the cells of action and cannot locate proteins or other biological molecules. The power of cell imaging is universally acknowledged. Earlier this fall, three microscope pioneers won the Nobel Prize in Chemistry this year for developing a super-resolution fluorescence microscope. Just like the progress of super-resolution microscopes, it can fundamentally change the way we look at the cellular world. However, whether it is a major milestone or a daily progress, we can save us time and resources on the bench. The new imaging technology is always well received and welcomed by the research community. To this end, BioTechniques' editor reviewed the progress of microscope technology over the past year and selected our favorite paper published in 2014. Our choice clearly shows that the increasing diversity of imaging methods is being applied to today's life science research.

1. "Two-color fluorescent in situ hybridization using chromogenic substrates in zebrafish," by Schumacher et al. (November 2014)

When it comes to the microscope, we were spoiled. Most of the confocal images we see come in a variety of colors and provide a range of data. However, for some technologies, these colors pay a price. For two-color fluorescence in situ hybridization (FISH), the cost is always high to detect weakly expressed transcripts and to monitor the signal intensity and background values ​​during the experiment. The research group of Saulius Sumanas at the Children's Hospital of Cincinnati further studied the possibility of using chromogenic substrates instead of traditional labeled probes for FISH. Combining NBT/BCIP and Vector Red, which have non-overlapping reflected wavelengths, the authors created a program to compare the long reactivity of alkaline phosphatase, the chromogenic monitoring of development reactions, and high-resolution fluorescence imaging. Zebrafish gene expression patterns. [literature]

2. "Robust and artifact-free mounting of tissue samples for atomic force microscopy," by Morgan et al. (September 2014)

Atomic force microscopy (AFM) is a technique used to study the physical properties of cells and tissues. One disadvantage of AFM is that it requires fixing the tissue sample before imaging. Fixation is usually done by glue or dry samples, both of which may produce artificial errors. To eliminate this possible source of error, Paul Russell and colleagues at the University of California, Davis, built a device that they called the Tissue Clamp Fixation Holder (SCIRT) to use to fix AFM samples. Using SCIRT, Russell's research team was able to process small samples, provide continuous hydration of the sample, eliminate glue and its associated artifacts, and even recover samples after AFM measurements. [literature]

3. "Multi-modality imaging of a murine mammary window chamber for breast cancer research," by Schafer et al. (July 2014)

Sometimes it is more cost-effective to use one or more techniques to image samples or specimens. Optical microscopy can provide information on cell-level details, such as magnetic resonance imaging (MRI), which can provide larger structures with high-resolution morphological information, such as tumor size and shape. In July of this year, Arthur Gmitro and colleagues at the University of Arizona in the United States introduced their new method in detail for microanimal tumor microenvironment imaging. Researchers used an implanted breast window to image the tumor environment with light microscopy and MRI and nuclear imaging. With the same breast window, the ability to focus on a single anatomical area with multiple imaging techniques can provide new insights into the relationship between breast cancer cells and tumor growth. [literature]

4. "Investigation of membrane protein-protein interactions using correlative FRET-PLA," by Ivanusic et al. (October 2014)

Not all new imaging techniques will produce bright, high-contrast color pictures and win microscope image competitions, but even an unattractive approach can still produce great information. Daniel Ivanusic and his colleagues in Berlin, Germany, published an example in October this year. Techniques such as fluorescence energy resonance transfer (FRET) and proximity ligation technology (PLA) can be used to monitor whether and when proteins interact. Ivanusic's team realized that combining the relevant FRET and PLA technologies may be able to detect membrane protein interactions better than using each technique individually. They found that in the study of protein interactions, this series of experiments can verify the robustness and reliability of the related FRET-PLA. [literature]

5. "Nuclear LC3-positive puncta in stressed cells do not represent autophagosomes," by Buckingham et al. (November 2014)

Finally, there are times when you need to remind you that seeing does not necessarily mean believing. In November, Charles Grose and his research team at the University of Iowa conducted an in-depth study of the recent observations of two research groups that studied cell nuclear LC3-positive spots in cells. The LC3 antibody is associated with the autophagosome, which should mean the nuclear localization of the autophagosome - what was previously thought to be non-existent. Grose and his team found that the observed staining was not specific to LC3, but was due to non-specific staining caused by a certain permeability and hybridization conditions.