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Single molecule localization microscopy for quantitative biology

Single-molecule localization microscopy (SMLM) generates super-resolution images by serially detecting individual fluorescent molecules. The power of SMLM, however, goes beyond images: biologically relevant information can be extracted from the mathematical relationships between the positions of the fluorophores in space and time. These analytical methods allow new routes to quantify spatial patterning, clustering, and colocalization at nanometer length scales.

Systems imaging to reveal the eukaryotic organelle interactome and oral microbiome structure and assembly

To fully understand the function of any biological system, it is often necessary to know how the different components of the system are arranged in space.  However, the ability to distinguish more than a few different components using fluorescence microscopy is severely limited.  My laboratory has employed spectral imaging and novel computational analyses to study the structure of two biological systems: the eukaryotic cell and the human oral microbial community.

Challenges of Establishing a new imaging facility: The floating core diaries

Core facilities are an essential resource in research institutes. They provide dedicated expertise and the instrumentation necessary for the development and improvement of science in academia. However, establishing a new microscopy core facility in a competitive environment like Harvard Medical School is not extent of challenges. In most cases it requires the creation of dedicated spaces, which are hard to come by and/or expensive to renovate, to host the instruments, and attracting users to obtain a solid user base is essential for the success of the core.

Serial block face imaging

We have recently installed a serial block face imaging system and have collected our first major data set. The project was a proof of concept to track the progress of pollen tubes towards specialist cells involved in plant reproduction. Pollen tubes target synergid cells and in particular the basal membrane of those cells which is known as the filiform apparatus.

(206) Integration and validation of RNA ISH, multispectral imaging and analysis protocols into a Core environment

Identification of biomarkers is a major goal of personalized medicine. Large transcriptome screens have identified new targets and molecular signatures for disease sub-types.  However, tissue spatial information, which fundamentally alters in vivo cell behavior and gene expression, is lost.  To understand spatial context and validate bulk tissue screens, most researchers rely exclusively on antibodies and immunostaining assays.

(209) Implementation of multiplex staining, imaging and analysis as a standardized service for researchers

Several modalities of multiplex immunofluorescence histology currently available require significant time and resources to implement. Many research laboratories develop questions benefiting from multiplex staining and analysis but do not have the human resources and/or equipment to perform the assay.