ABSTRACT: KIDNIT (Kidney Imaging: Developing Novel and Innovative Tools) is the Pittsburgh Center for Kidney Research’s Resource Development Core. Working in collaboration with kidney investigators and the Center’s Physiology and Model Systems Core, KIDNIT will build and adapt cutting-edge light, super-resolution, and electron microscopic imaging modalities and technologies to address open questions related to kidney function and dysfunction. The broad and long-range goals of KIDNIT are as follows: Aim 1 is to nurture a dynamic incubator space for imaging tool development. KIDNIT will capitalize on the resources provided by the Center for Biological Imaging (CBI), one of the largest imaging facilities in the country. The CBI has a dedicated expert faculty, a large cohort of research technicians, and a vast array of imaging equipment including over 40 contemporary widefield, confocal, two-photon, TIRF, light-sheet, super-resolution, and electron microscopes. Aim 2 is to develop new and custom-made imaging resources, tools, and technologies specifically to augment and advance kidney research. KIDNIT will leverage the CBI’s faculty expertise to develop, adapt, and implement new imaging tools for kidney researchers. Example projects in development include novel clearing and 3D imaging protocols to reveal and quantify kidney architecture and function at the level of the entire organ; analyzing collagen organization and structure in normal and fibrotic tissues using a recently developed variant of polarized light microscopy (instant polarized light microscopy; iPOL); development of new protocols that employ fluorescent lifetime imaging and fluorescent energy transfer to rapidly assess kidney cell function and responses to extracellular cues; use of high throughput platforms to perform large-scale screening of organoid form and function; and development of novel microfluidic chambers. Mechanisms are in place to ensure training and information transfer. Aim 3 is focused on ensuring robust validation and quality control of new resources. An example is a validated antibody collection that can be used to assess differentiation, injury, and repair status of kidney cells and tissues. Aim 4 is to collaborate with the Consortium to prioritize, suspend, or implement new and improved imaging resources. Working in collaboration with the Consortium, KIDNIT will replace obsolete technologies by leveraging its ties with industry leaders, along with a strong track record of S10 funding, to offer the most recent developments in microscopy, specifically to advance the research of our investigator collective. Impact: KIDNIT will advance kidney research by providing Center and Consortium investigators with novel and bespoke imaging tools that will provide critical information about the location, dynamics, amounts, and function of kidney-associated molecules, organelles, cells, and tissues in normal and disease states.