The mission of the Twinbrook Imaging Facility is to provide the necessary instrumentation and technical support to allow researchers to acquire and analyze high-resolution images of living cells. Under Dr. Brzostowski,’s leadership, the facility provides training to fellows, students, and courageous PIs. Dr. Brzostowski is a strong believer in communication amongst users to foment academic growth and solve common problems. It is the philosophy of the facility to maintain fluidity in the open design of microscopes to meet the changing needs of the investigators using the systems.
Integral to the imaging facility is the computer workstation corral for data analysis and its human keeper Dr. Javier Manzella-Lapeira, located in Fishers Lane, room 4S06. Dr. Manzella-Lapeira manages three high-end PCs and one Mac workstation. Users have access to unlimited data storage that is backed up daily and access to the analysis software listed below. While the corral is a quiet space for work and study, it also promotes conversation among users to discuss data and technical issues. Utmost to the facility, are the excellent programming and imaging skills of Dr. Manzella-Lapeira. Dr. Manzella-Lapeira has written a variety Matlab code tailored to tackle a number of complicated data analysis challenges. Current algorithms are freely available or can be modified to apply to new problems in a collaborative manner upon request.
- Zeiss ZEN
- Imaris (supported through NIAID core imaging facility)
- Huygens 3D deconvolution
- Dynamic Information Architecture System (DIAS) (cell tracing/tracking)
The facility supports a number of light microscopy techniques to image live cells that include laser scanning confocal microscopy, spectral imaging, spinning disk confocal microscopy, FRET, total internal reflection fluorescence microscopy, single molecule traclomg, super-resolution STORM imaging, Airyscan super-resolution imaging, fluorescence lifetime imaging, fluorescence correlation spectroscopy, and intravital imaging. While we may have a penchant for the exotic, the facility whole-heartedly welcomes routine imaging of fixed samples and supplies a host of nail polish hues to adorn user’s slides.
Imaging facility group members have written several book chapters that detail methods for acquiring and analyzing FRET and single particle data using TIRFM and FRET by confocal microscopy.
Hae Won S, Brzostowski J. Time-lapse Forster resonance energy transfer imaging by confocal laser scanning microscopy for analyzing dynamic molecular interactions in the plasma membrane of B cells. Methods in Mol Biol. [ePub ahead of print].
Xu X, Yun M, Wen X, Brzostowski J, Quan W, Wang QJ, Jin T. Quantitative monitoring of tempo-spatial activation of Ras and PKD1 using confocal fluorescent microscopy. Methods Mol Biol. 2016;1407:307-23.
Davey A, Liu W, Sohn HW, Brzostowski JA, Pierce SK. Understanding the initiation of B cell signaling through live cell imaging(link is external).Methods Enzymol. 2012;506:265-90.
Sohn HW, Tolar P, Brzostowski J, Pierce SK. A method for analyzing protein-protein interactions in the plasma membrane of live B cells by fluorescence resonance energy transfer imaging as acquired by total internal reflection fluorescence microscopy.(link is external) Methods Mol Biol. 2010;591:159-83.
Xu X, Brzostowski JA, Jin T. Monitoring dynamic GPCR signaling events using fluorescence microscopy, FRET imaging, and single-molecule imaging.(link is external) Methods Mol Biol. 2009;571:371-83.
Brzostowski JA, Meckel T, Hong J, Chen A, Jin T. Imaging protein-protein interactions by Förster resonance energy transfer (FRET) microscopy in live cells.(link is external) Curr Protoc Protein Sci. 2009 Apr;Chapter 19:Unit19.5.