Light Microscopy core for NIDDK

National Institute Of Diabetes And Digestive And Kidney Diseases

Investigators

Abstract

The mission of the National Institutes of Health (NIH) is to seek fundamental knowledge about the nature and behavior of living systems and to apply that knowledge to enhance health, lengthen life, and reduce illness and disability. As part of the NIH, the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) Intramural Research Program (IRP) conducts biomedical research and training related to diabetes mellitus; endocrine, bone, and metabolic diseases; digestive diseases, including liver diseases and nutritional disorders; and kidney, urologic, and hematologic diseases. Intramural research is conducted in the Institute's laboratories and clinical facilities in Bethesda, Maryland and in Phoenix, Arizona. There are currently 18 Laboratories and Branches (i.e. 18 departments with different focus areas) in the NIDDK IRP. Most if not all of these areas of research, either occasionally or on an ongoing basis, require light microscopy (LM) methods to qualitatively complement other assays, or as the basis of a quantitative assay. There are three main categories of LM. Conventional widefield, transmitted light techniques, such as darkfield or differential interference contrast (DIC), are still useful because of their innate ability to show cellular morphology. Second, an old standby, immunohistochemistry (IHC)-- using absorptive probes to colorize tissue in transmitted light-- has been used for decades (e.g. in pathology), but recent advances with new probes and color deconvolution have allowed the field to grow, although requiring more finesse when being quantitated. The third and most common category of LM is characterized by the use of fluorescent probes targeting biomolecules (specific proteins, DNA, etc) of interest, making them glow with different colors on a black background. Such fluorescent dyes can even be used to monitor subcellular events in real-time. There are many sub-categories of fluorescence microscopy, e.g. confocal, super-resolution, and TIRF (Total Internal Reflectance-induced Fluorescence). Each of these involve specialized technique in sample prep, image acquisition, and/or image analysis. Some NIDDK labs might be comfortable without advanced help on one of more of the techniques described above, and might also be able to set aside a budget for equipment they would use regularly; but to fill any gaps, the NIDDK Advanced Light Microscopy & Image Analysis Core (ALMIAC) remains a reliable resource for cutting-edge light microscopy equipment, software, and expertise. For the FY2023 budget year (October 2022 - September 2023), there were >60 researchers using ALMIAC resources, representing at least 33 (out of 96, 30%) "Sections" (individual labs), from at least 11 (out of 18, or 60%) Laboratories and Branches. Out of the 60+ individual researchers helped, there were at least 5 NIDDK publications during FY 2023 (and several more in preprint stage as of 8/31/2023, not listed below) that used images acquired and/or analyzed with the help of ALMIAC resources: --Cheng-Chao Lin, Luis F Menezes, Jiahe Qiu, Elisabeth Pearson, Fang Zhou, Yu Ishimoto, D Eric Anderson, Gregory G Germino. In vivo Polycystin-1 interactome using a novel Pkd1 knock-in mouse model. PLoS One. 2023 Aug 4;18(8):e0289778. PMID:37540694 PMCID:PMC10403143 DOI:10.1371/journal.pone.0289778. --Daniel Stoyko, Timothy O, Adrianna Hernandez, Parthena Konstantinidou, Qingcai Meng, Astrid D Haase. CRISPR-Cas9 Genome Editing and Rapid Selection of Cell Pools. Curr Protoc. 2022 Dec;2(12):e624. PMID:36546759 PMCID:PMC9793982(available on 2023-12-01) DOI:10.1002/cpz1.624 --Naoko Tsuji, Takayuki Tsuji, Tetsushi Yamashita, Naoki Hayase, Xuzhen Hu, Peter St Yuen, Robert A Star. BAM15 treats mouse sepsis and kidney injury, linking mortality, mitochondrial DNA, tubule damage, and neutrophils. J Clin Invest. 2023 Apr 3;133(7):e152401. PMID:36757801 PMCID:PMC10065071 DOI:10.1172/JCI152401. --Lihui Wang, Yue Xu, Sijung Yun, Quan Yuan, Prasanna Satpute-Krishnan, Yihong Ye. SAYSD1 senses UFMylated ribosome to safeguard co-translational protein translocation at the endoplasmic reticulum. Cell Rep. 2023 Jan 31;42(1):112028. PMID:36848233 PMCID:PMC10010011 DOI:10.1016/j.celrep.2023.112028 --Yu Ishimoto, Luis F Menezes, Fang Zhou, Teruhiko Yoshida, Taishi Komori, Jiahe Qiu, Marian F Young, Huiyan Lu, Svetlana Potapova, Patricia Outeda, Terry Watnick, Gregory G Germino. A novel ARPKD mouse model with near-complete deletion of the Polycystic Kidney and Hepatic Disease 1 (Pkhd1) genomic locus presents with multiple phenotypes but not renal cysts. Kidney Int. 2023 Sep;104(3):611-616. PMID:37419448 DOI:10.1016/j.kint.2023.05.027 In addition to specific research projects, several areas of improvement in capability and workflow include: 1) Addition of 2 new microscopes into the ALMIAC: an OMX SR (Super-Resolution via Structured Illumination) microscope; and a Zeiss spinning disk confocal for those NIDDK researchers in Building 50. 2) Upgrades to several ALMIAC microscopes; and helping with upgrades to NIDDK microscopes outside the ALMIAC. 3) Formation of a Core Advisory Group for the ALMIAC, who will meet in Fall 2023, and subsequently 1-2x/year.