Ragon Services - The Ragon Institute of MGH, MIT and Harvard


	Ragon Services
	Flow Cytometry Flow Cytometry is a technique that allows researchers to identify subpopulations within a heterogeneous mixture of cell types by measuring up to 20 light scattering and fluorescent parameters at a time, at rates as high as twenty thousand cells per second. Cell sorters are specialized flow cytometers that are then capable of separating out specific populations of interest from that mixture, collecting up to 4 different subtypes at a time into individual tubes, for either culturing or for further analysis through techniques such as gene expression arrays and western blotting. The Ragon Institute contains 5 analysis instruments capable of measuring 2 scatter parameters and from 4 to 18 colors, as well as a 12 color cell sorter. Visit Flow Cytometry on Ragon Institute Services for more information.

	Microscopy Confocal microscopy utilizes a pinhole in plane confocal to the sample which, when properly adjusted, can exclude scattered light from above and below the focal plane of the specimen. In this way, one is able to obtain very thin optical sections using the confocal microscope. This is also accomplished through the use of multiple wavelengths of laser light. The Ragon Institute Imaging Core utilizes a Zeiss LSM 510 confocal microscope that can image four different fluorescent signals simultaneously, and up to eight discrete signals sequentially, thereby taking full advantage of multichannel imaging. Images from multiple channels can thereby be combined to reveal precise localization of specific molecules or structures within tissues or cells. An additional benefit of confocal microscopy is that serial optical sections can be obtained in the vertical axis of the sample so that one can effectively “slice through” the sample or cell and obtain a three-dimensional reconstruction of that sample or cell based solely on the fluorescent signals. Since the confocal microscope uses photomultiplier tubes to record fluorescence intensity from each part of the sample imaged, intensity of multiple channels (fluorophores) can be quantified both in space and time. Since the confocal microscope uses lasers to scan the sample, obtaining high resolution images takes some time (on the order of one or several minutes). Therefore, the Imaging Core also is equipped with a Zeiss AxioObserver fluorescence microscope configured with high-speed filter wheels and shutters for rapid, live-cell multichannel time-lapse fluorescence imaging. Visit Microscopy on Ragon Institute Services for more information.

	Humanized Mouse Model of HIV Infection Despite great effort, an ideal animal model in which to study HIV pathogenesis and test vaccine efficacy remains elusive. Recently, we and others have developed a markedly improved humanized mouse model of HIV by transplanting human CD34+ stem cells and autologous human thymic grafts into immunodeficient mice In this model, we have achieved robust repopulation of mouse lymphoid tissues with human immune cells, and have generated robust anti-HIV cellular and humoral immune responses in these humanized mice. We believe this improved humanized mouse model will allow us to study questions regarding the biology of HIV-1 not readily approachable through human studies. The Ragon Institute Humanized Mouse Program was established in order to further develop this model, and to make it available to the wider Ragon research community. Highly experienced personnel produce these animals and perform experiments in collaboration with Ragon investigators at very reasonable cost. Visit Humanized Mouse on Ragon Institute Services for more information.

	Virology As part of the Ragon Institute's overall goal to study the pathogenesis of HIV and develop effective vaccine approaches, the Virology Platform provides critical infrastructure to provide a centralized facility for preparation, titreing, and storage of HIV stocks for immunological and virological studies. We also offer viral sequencing services and the evaluation of viral replication capacity of HIV strains and constructs. Visit Virology on Ragon Institute Services for more information.

	Tissue The Ragon Tissue Platform has been established to provide the necessary infrastructure to conduct patient-based research investigating tissue specific immune responses against HIV. We seek to significantly reduce the major obstacle to initiating tissue based studies by providing access to human tissue specimens to assist in studies of HIV pathogenesis and tissue-specific immune responses against HIV. Visit Tissue on Ragon Institute Services for more information.

	Biostatistics The Biostatistics Core serves the needs of the HIV/AIDS researchers within the Ragon Institute and its affiliates. In particular, members of the Biostatistics Core provide expertise in the planning, conduct and analysis of research with the goal of enhancing the scientific quality of HIV-related research at the institute. The primary objective of the core is to ensure that studies are well designed, correctly analyzed, clearly presented, and correctly interpreted. Visit Biostatistics on Ragon Institute Services for more information.