Highly Multiplexed Optically Sectioned Spectroscopic Imaging in a Programmable Array Microscope

Hanley, Quentin S.; Jovin, Thomas M.

doi:10.1366/0003702011953171

Cited by 11 publications

(17 citation statements)

References 32 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…PAMs have also been used in nonsectioning configurations for spatial encoding in two-and three-dimensional imaging spectroscopic applications (24), optically sectioning imaging spectroscopy (21,25), and a widefield system for measuring spectrally resolved FLIM (26).…”

mentioning

confidence: 99%

Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM)

et al. 2005

Self Cite

View full text Add to dashboard Cite

Background: The programmable array microscopes (PAMs) are a family of instruments incorporating arbitrary control of the patterns of illumination and/or detection. The PAM can be used in sectioning and nonsectioning modes, thereby constituting a useful platform for fluorescence lifetime imaging. Methods and Results: We used a PAM for acquisition of optically sectioned and widefield fluorescence lifetime images, in which contrast was increased predominantly by suppressing out-of-focus light contributions. We simu-

show abstract

mentioning

confidence: 99%

Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM)

et al. 2005

Self Cite

View full text Add to dashboard Cite

show abstract

“…The defining feature of a PAM is the presence of a spatial light modulator (SLM) in the image plane, where it is used to define a pattern of conjugate illumination and detection. The family of PAMs include optical sectioning systems operating in fluorescence and reflection modes, as well as spectroscopic imaging systems [5][6][7][8][11][12][13]. In addition, the programmable SLM allows the implementation of flexible photoactivation and photodepletion strategies [14].…”

Section: The Programmable Array Microscope (Pam)mentioning

confidence: 99%

Biological applications of an LCoS-based programmable array microscope (PAM)

et al. 2007

Self Cite

View full text Add to dashboard Cite

We report on a new generation, commercial prototype of a programmable array optical sectioning fluorescence microscope (PAM) for rapid, light efficient 3D imaging of living specimens. The stand-alone module, including light source(s) and detector(s), features an innovative optical design and a ferroelectric liquid-crystal-on-silicon (LCoS) spatial light modulator (SLM) instead of the DMD used in the original PAM design. The LCoS PAM (developed in collaboration with Cairn Research, Ltd.) can be attached to a port of a(ny) unmodified fluorescence microscope. The prototype system currently operated at the Max Planck Institute incorporates a 6-position high-intensity LED illuminator, modulated laser and lamp light sources, and an Andor iXon emCCD camera. The module is mounted on an Olympus IX71 inverted microscope with 60-150X objectives with a Prior Scientific x,y, and z high resolution scanning stages. Further enhancements recently include: (i) point-and line-wise spectral resolution and (ii) lifetime imaging (FLIM) in the frequency domain. Multiphoton operation and other nonlinear techniques should be feasible. The capabilities of the PAM are illustrated by several examples demonstrating single molecule as well as lifetime imaging in live cells, and the unique capability to perform photoconversion with arbitrary patterns and high spatial resolution. Using quantum dot coupled ligands we show real-time binding and subsequent trafficking of individual ligand-growth factor receptor complexes on and in live cells with a temporal resolution and sensitivity exceeding those of conventional CLSM systems. The combined use of a blue laser and parallel LED or visible laser sources permits photoactivation and rapid kinetic analysis of cellular processes probed by photoswitchable visible fluorescent proteins such as DRONPA.

show abstract

“…The evolution of PAMs has encompassed a variety of imaging concepts and technical realizations: confocal microscopy (1-9), imaging spectroscopy (10-12), deconvolution of conjugate and nonconjugate images (9), optical sectioning integrated with imaging spectroscopy (13), and lifetime imaging combined in hybrid systems with optical sectioning (14) or imaging spectroscopy (15). In these implementations, micromirror devices (1-9,16), nematic liquid crystals (10), and ferroelectric liquid crystals (7) have served as SLMs for control of the patterns of illumination and detection.…”

Section: Analytical Cytologymentioning

confidence: 99%

Selective photoreactions in a programmable array microscope (PAM): Photoinitiated polymerization, photodecaging, and photochromic conversion

et al. 2005

Self Cite

View full text Add to dashboard Cite

BackgroundInnovative thinking and experimentation were the hallmarks of Mack Fulwyler's approach to research. This report summarizes some of the ideas and their early realizations that he pursued in the field of imaging cytometry, work that was not published before his untimely death, although he composed the initial draft of this report.MethodsIncluded are related experiments implemented in the programmable array microscope (PAM) devised for patterned illumination and detection, the instrument that Mack Fulwyler employed during a sabbatical leave in Göttingen in 1998. Despite being the originator of instrumentation for flow cytometry and sorting, Mack Fulwyler was intensely interested in imaging systems, recognizing their ability to resolve cellular details obscured by the whole cell signals generally acquired in flow. At one point, these interests merged with those of two other authors (I.T.Y. and T.M.J.), leading to the Image Cytometry and Sorting (ICAS) strategy and project. A major goal was uncomplicated rare cell detection and isolation using a sequential process of cellular labeling via suitable probes, whole field imaging, and selective area‐restricted photoinduced reactions designed to encapsulate and/or chemically or physically tag cells in a manner permitting subsequent fractionation by bulk techniques.Results and ConclusionThis publication features photoinduced polymerization, photodecaging, photoactivation, and photochromic conversion reactions carried out by Fulwyler and/or the other authors with the PAM, employing operator designated patterns and locations in various samples. Photopolymerization of polyethylene glycol‐diacrylate to a gel‐like structure allowing the specific selection of objects (cells) for further analysis and processing techniques was the approach explored personally by Mack Fulwyler in relation to the ICAS concept. © 2005 International Society for Analytical Cytology

show abstract

Highly Multiplexed Optically Sectioned Spectroscopic Imaging in a Programmable Array Microscope

Cited by 11 publications

References 32 publications

Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM)

Fluorescence lifetime imaging in an optically sectioning programmable array microscope (PAM)

Biological applications of an LCoS-based programmable array microscope (PAM)

Selective photoreactions in a programmable array microscope (PAM): Photoinitiated polymerization, photodecaging, and photochromic conversion

Contact Info

Product

Resources

About