Wang, Yao et al. (2021) International Worm Meeting "A simple and inexpensive add-on enables confocal imaging capacity on a widefield microscope"

Chung, Samuel, Grooms, Noa, Civale, Sabrina, Wang, Yao

[International Worm Meeting, 2021]

Widefield microscopes are commonly used in many biological laboratories. However, their inability to reject scattered or out-of-focus light often produces images with obscured thin or dim structures when there are some bright structures nearby. Scanning microscopes like confocal microscopes and multiphoton microscopes can solve this problem, but they are comparatively slower and much more expensive. We developed an inexpensive way to empower confocal imaging capacity on a widefield microscope, by inserting a spatial light modulator (SLM) into the field stop of the widefield microscope and customizing the illumination pattern and acquisition methods. We assessed the performance of this SLM-inserted setup by comparing images taken at our widefield microscope, our widefield microscope with the SLM-inserted setup, and a commercial confocal microscope. While a widefield microscope showed no sectioning capability, our SLM-inserted setup showed 0.85 ± 0.04 mum and the commercial confocal showed 0.68 ± 0.04 mum optical sectioning capability. Additionally comparing images of the FLP neuron and the tightly bundled amphid neurons in C. elegans taken by the widefield, SLM-inserted setup, and confocal microscopes, we confirmed that the SLM-inserted setup greatly reduces haze from the bright cell body, allowing visualization of dim axons and dendrites nearby. Our SLM-inserted setup identified 96% of the dim neuronal fibers seen in confocal images while the widefield microscope only identified 50% of the same fibers. Our SLM-inserted setup represents a very simple (2-component) and inexpensive (<$600) approach to enable confocal capacity on a widefield microscope. This SLM-inserted setup can be broadly employed by labs that are using widefield microscopes, with minimum expense and modification.

Wang, Yao et al. (2019) International Worm Meeting "High-throughput Microscopy Platform toward a Roadmap for Accelerating Vertebrate Neuroregeneration Research."

Jaklitsch, Erik, Chung, Samuel, Wang, Yao

[International Worm Meeting, 2019]

Statement of Purpose Lesion conditioning to a neuron activates cellular mechanisms to enhance neuro-regeneration. Our prior study established the roundworm C. elegans as a powerful model for the rapid discovery of genes underlying lesion conditioning, which is impractical in vertebrates (Chung, S. H., et al. (2016).) Even so, a complete search for regeneration genes requires surgery and reimaging of a fluorescent neuron in >30,000 C. elegans, which represents significant efforts for the gene screening. A high-throughput surgery and reimaging platform, or specifically, a new integrated microscope platform for rapid immobilization, imaging, and optical surgery of C. elegans, would enable this gene search. Method Used Achievement of this goal requires the acceleration of two time-consuming steps: animal immobilization and precise neuronal microscopy. This can be accomplished through two specific goals: 1. Develop a microscope stage to readily immobilize many animals for in vivo imaging. 2. Enable robust automation of microscopy by improving neuron-background contrast. For Goal 1, C. elegans will be immobilized by thermoelectrically cooling them directly on their cultivation plates, instead of mounting them to slides. For Goal 2, patterned illumination approaches will enhance contrast by reducing the excitation power that illuminates extremely bright cell bodies. This patterned illumination will reduce the bright haze around the cell body from obscuring the axon and dendrites, and exposes the axon and dendrites near the cell body. Summary of Result We finished the thermal simulation of the cooling stage and tested thermoelectric performance under experimental conditions, which confirmed the feasibility of immobilizing worms on their cultivation plates. We invented a new coarse segmentation algorithm and patterned the illumination by employing a spatial light modulator, which validates our contrast improvement approaches. The accomplishment of our goals will pave the path to the high-throughput microscope platform for C. elegans research, which can be used in numerous neuroscience labs worldwide. This platform also enables our regeneration screen. Affiliations: Funding provided by Northeastern University

(2008) Sci China C Life Sci "Retraction: Wang, Yao et al. The flhDC gene affects motility and biofilm formation in Yersinia pseudotuberculosis. Sci China Ser C-Life Sci, 2007, 50(6): 814-821."

[Sci China C Life Sci, 2008]

(2009) Sci China C Life Sci "Retraction: Wang Yao, et al. The flhDC gene affects motility and biofilm formation in Yersinia pseudotuberculosis. Sci China Ser C-Life Sci, 2007, 50(6): 814-821."

[Sci China C Life Sci, 2009]

Infarinato N (2019) J Cell Biol "Xiaochen Wang: Building up our understanding of breaking down."

Infarinato N

[J Cell Biol, 2019]

Wang studies lysosomal degradation pathways using <i>C. elegans</i> as a model system.

Wang JT et al. (2014) Curr Biol "P granules."

Seydoux G, Wang JT

[Curr Biol, 2014]

Wang and Seydoux discuss the functional importance of P granules - the germline-specific RNA granules of C. elegans.

Wang X et al. (2010) Zhongguo Zhong Yao Za Zhi "[Effect of Ligusticum chuanxiong extract on lifespan of Caenorhabditis elegans and its underlying molecular mechanisms]"

Wang D, Niu X, Li L, Wang X

[Zhongguo Zhong Yao Za Zhi, 2010]

OBJECTIVE: To explore the effect of Ligusticam chuanxiong extract (CXE) on lifespan of Caenorhabditis elegans and investigate its underlyirig molecular mechanisms. METHOD: The lifespan assay was carried out on animals grouped into blank control group and CXE groups with concentration from low to high: 12.5, 25, 50, 100 mg x L(-1) by examining the effect of CXE on mean lifespan and maximum lifespan of C. elegans. According to the result of lifespan assay, we cultured the animals with the optimal concentration of CXE for 10 days, and tested the expression change of aging-related genes between the control and CXE group by realtime RT-PCR (qRT-PCR). RESULT: Compared with the control, 25, 50, 100 mg x L(-1) CXE all significantly extended the mean lifespan (15.7%, 9.1%, 6.2% respectively) and the maximum lifespan (15.0%, 6.8%, 6.6% respectively) of C. elegans. After treatment with 25 mg x L(-1) CXE the expression of hsp-70, skn-1 were obviously up-regulated while the expression of akt-2, tub-1 were significantly down-regulated. CONCLUSION: CXE significantly extend the lifespan of C. elegans, and the underlying molecular mechanism is related with genes of Insulin/IGF-1 signaling pathway and dietary restriction system.

Wang X et al. (2010) Zhongguo Zhong Yao Za Zhi "[Study of molecular mechanism of anti-Parkinson's disease traditional Chinese medicine using model of Caenorhabditis elegans]"

Gao H, Wang D, Wang X

[Zhongguo Zhong Yao Za Zhi, 2010]

Parkinson's disease (PD) is a neurodegenerative disorder with a complex, multifactorial aetiology. The brains of patients affected with PD are characterized by a loss of neurons in dopamine neurons in the substantia nigra, decreasing of dopamine secretion, and the deposition of Lewy bodies (LBs) in the cytoplasm of remaining neurons. In China the data show that the incidence of Parkinson's disease increases at least 20 times in recent 20 years, and it makes things worse for the aging society. Developing good anti-PD drugs to improve the patient's quality of life is particularly important. The treatment of PD using traditional Chinese medicine (TCM) has made remarkable effect, while the the molecular mechanisms of it is still not known, while elucidating the molecular mechanism of TCM is the base of better understanding its function. Using genetically modified PD model of Caenorhabditis elegans, which is suitable for molecular mechanism study, to explore the interference mechanism of TCM to PD might be an effective way. This review briefly introduces the research progress on molecular mechanism of PD, and then discusses the idea of using C. elegans to study molecular mechanism of TCM intervention to PD.

Wang X et al. (2010) Zhongguo Zhong Yao Za Zhi "[Expression changes of age-related genes in different aging stages of Caenorhabiditis elegans and the regulating effects of Chuanxiong extract]."

Wang D, Li L, Niu X, Wang X

[Zhongguo Zhong Yao Za Zhi, 2010]

OBJECTIVE: To explore the expression changes of age-related genes in different stages of aging and the regulating effects of Chuanxiong extract on it. METHOD: According to the different stages of aging, the experiments were tested at two time points of 2 d and 6 d. Using realtime RT-PCR (qRT-PCR) to test the expression change of aging-related genes among the groups. RESULT: Compared with the 2 d control group, the expression of age-1, daf-2, let-363 were up-regulated in the 6 d control group (P &lt; 0.05) while the expression of ins-18, let-60, sir-2.1, sod-3 were down-regulated (P &lt; 0.05). Compared with the 2 d administration group, the expression of age-1, daf-2, let-363 were significantly up-regulated (P &lt; 0.01) in the 6 d administration group after treated with CXE while the expression of ins-18, let-60, sir-2.1, sod-3 were significantly down-regulated (P &lt; 0.01). CONCLUSION: In the progress of aging, the expression of age-1, daf-2, let-363 increased, functioning as aging-promoting genes; while the expression of ins-18, let-60, sir-2.1, sod-3 decreased, functioning as longevity genes; CXE extended the lifespan through inhibiting the expression of these aging-promoting genes and increasing the expression of longevity genes, which would be the molecular mechaniSm of anti-aging of traditional Chinese medicine that can promote Qi and activate blood.

Munro E (2017) Dev Cell "Protein Clustering Shapes Polarity Protein Gradients."

Munro E

[Dev Cell, 2017]

In this issue of Developmental Cell, Dickinson etal. (2017) and Rodriguez etal. (2017), along with Wang etal. (2017) in Nature Cell Biology, show how PAR protein oligomerization can dynamically couple protein diffusion and transport by cortical flow to control kinase activity gradients and polarity in the C.elegans zygote.