Automated detection of rare‐event pathogens through time‐gated luminescence scanning microscopy

Lu, Yiqing; Jin, Dong-Yan; Leif, Robert C.; Deng, Wei; Piper, James A.; Yuan, Jingli; Duan, Yusheng; Huo, Yujing

doi:10.1002/cyto.a.21045

Cited by 22 publications

(31 citation statements)

References 26 publications

(26 reference statements)

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“…The light emitted at a large solid angle ([458) is collected by a UV-fused silica plano-convex lens (f 5 35.mm; Ø51 inch, Thorlabs LA4052UV), which is then redirected by a dichroic mirror (365 DCLP BS, http://www.chroma.com/) into the UV objective. Though mechanical chopper gating has been reported with slow switching times of 100-200 ls (14,32-36), we employed a new compact fast chopper (C995 Optical Chopper, Terahertz Technologies, NY; 167 rotating cycles per second) (30,37) and a new optics configuration to achieve up to 11 ls switching time at 2.5-kHz repetition rate with a transparent/blocking duty ratio of 75%:25% (30). This duty ratio is achieved by laser micromachining of every second blades from original 30 blades down to 15 blades.…”

Section: Methodsmentioning

confidence: 99%

Demonstration of true‐color high‐contrast microorganism imaging for terbium bioprobes

Jin

2011

Cytometry Pt A

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Lanthanide bioprobes offer a number of novel advantages for advanced cytometry, including the microsecond luminescence lifetime, sharp spectral emission, and large stokes shift. However, to date, only the europium-based bioprobes have been broadly studied for time-gated luminescence cell imaging, though a wide range of efficient terbium bioprobes have been synthesized and some of them are commercially available. We analyze that the bottleneck problem was due to the lack of an efficient microscope with pulsed excitation at wavelengths of 300-330 nm. We investigate a recently available 315 nm ultraviolet (UV) light emitting diode to excite an epifluorescence microscope. Substituting a commercial UV objective (403), the 315 nm light efficiently delivered the excitation light onto the uncovered specimen. A novel pinhole-assisted optical chopper unit was attached behind the eyepiece for direct lifetime-gating to permit visual inspection of background-free images. We demonstrate the use of a commercial terbium complex for high-contrast imaging of an environmental pathogenic microorganism, Cryptosporidium parvum. As a result of effective autofluorescence suppression by a factor of 61.85 in the time domain, we achieved an enhanced signal-to-background ratio of 14.43. This type of time-gating optics is easily adaptable to the use of routine epifluorescence microscopes, which provides an opportunity for high-contrast imaging using multiplexed lanthanide bioprobes. ' International Society for Advancement of CytometryKey terms time-gated luminescence; bioimaging; terbium; UV LED; Cryptosporidium; high-contrast EPIFLUORESCENCE microscopes are routinely found in both research and diagnostic laboratories. From the practical point of view, cytometrists expect a microscope to be low cost, compact, easy-to-operate at high sensitivity, including the capacity to discriminate against nontarget substance, detect multiple species at once, and have high throughput. Time-domain discrimination of targeted microorganisms using long-lifetime lanthanide bioprobes shows promise in meeting these demands. This is due to the ease of time-gating of probes that have lifetimes of several hundreds of microseconds, which eliminates the autofluorescent background of several nanoseconds (1). In practice, however, there are several bottleneck problems challenging both the instrumentation and bioprobe developments. To date, only the europiumbased bioprobes (excited at 330-420 nm, emitting 615 nm at $10 nm bandwidth) have been broadly studied for time-gated luminescence cell imaging (1-15). This is due to the fact that other rare-earth ion complexes are either less efficient (16) [only europium and terbium complexes are less sensitive to vibrational quenching by energy transfer to OH, NH, or CH oscillators (15)] or require pulsed ultraviolet (UV) excitation below 330 nm (17). Another reason is the lack of near-infrared detection cameras. In fact, some cameras have included near-infrared blocking filters. For example, many efficient terbium-base...

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Section: Methodsmentioning

confidence: 99%

Demonstration of true‐color high‐contrast microorganism imaging for terbium bioprobes

Jin

2011

Cytometry Pt A

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“…The principles were described previously [1,4]. In this work, the major difference in system setup has been focused on improved scanning speed and accuracy.…”

Section: System Arrangementmentioning

confidence: 99%

“…Time-gated luminescence (TGL) technique is drawing increasingly more attentions in the analytical fields of microbiology and molecular biology, medical diagnostics, health and environmental researches as well as applications [1][2][3][4]. It takes advantage of long-lived luminescence from special substances, such as lanthanides exhibiting emission lifetime of >100 ȝs, which overwhelms autofluorescence, whose lifetime is usually < 0.1 ȝs.…”

Section: Introductionmentioning

confidence: 99%

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Cytometric investigation of rare-events featuring time-gated detection and high-speed stage scanning

Piper

Huo

et al. 2011

2011 International Quantum Electronics Conference (IQEC) and Conference on Lasers and Electro-Optics (CLEO) Pacific Rim Incorpo

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“…One solution to this problem includes the use of lanthanide luminescent materials exhibiting long lifetimes and/or photon upconversion properties, which are highly useful as either high-contrast molecular probes for direct labelling [24][25][26][27][28][29][30] or microsphere-based suspension arrays for high throughput assays [31][32][33] .…”

Section: Introductionmentioning

confidence: 99%

High-Precision Pinpointing of Luminescent Targets in Encoder-Assisted Scanning Microscopy Allowing High-Speed Quantitative Analysis

Zheng

Zhao

et al. 2015

Anal. Chem.

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The new agreement specifically addresses what authors can do with different versions of their manuscript -e.g. use in theses and collections, teaching and training, conference presentations, sharing with colleagues, and posting on websites and repositories. The terms under which these uses can occur are clearly identified to prevent misunderstandings that could jeopardize final publication of a manuscript (Section II, Permitted Uses by Authors). Easy Reference User Guide Posting Submitted Works on Websites and Repositories:A digital file of the unedited manuscript version of a Submitted Work may be made publicly available on websites or repositories (e.g. the Author's personal website, preprint servers, university networks or primary employer's institutional websites, third party institutional or subject-based repositories, and conference websites that feature presentations by the Author(s) based on the Submitted Work) under the following conditions:• The posting must be for non-commercial purposes and not violate the ACS' "Ethical Guidelines to Publication of Chemical Research" (see http://pubs.acs.org/ethics).• If the Submitted Work is accepted for publication in an ACS journal, then the following notice should be included at the time of posting, or the posting amended as appropriate: "This document is the unedited Author's version of a Submitted Work that was subsequently accepted for publication in [JournalTitle], copyright © American Chemical Society after peer review. To access the final edited and published work see [insert ACS Articles on Request author-directed link to Published Work, see http://pubs.acs.org/page/policy/articlesonrequest/index.html]." Note: It is the responsibility of the Author(s) to confirm with the appropriate ACS journal editor that the timing of the posting of the Submitted Work does not conflict with journal prior publication/embargo policies (see http://pubs.acs.org/page/policy/prior/index.html)If any prospective posting of the Submitted Work, whether voluntary or mandated by the Author(s)' funding agency, primary employer, or, in the case of Author(s) employed in academia, university administration, would violate any of the above conditions, the Submitted Work may not be posted. In these cases, Author(s) may either sponsor the immediate public availability of the final Published Work through participation in the feebased ACS AuthorChoice program (for information about this program see http://pubs.acs.org/page/policy/authorchoice/index.html) or, if applicable, seek a waiver from the relevant institutional policy. July, 2016 ABSTRACTCompared with routine microscopy imaging of a few analytes at a time, rapid scanning through the whole sample area of a microscope slide to locate every single target object offers many advantages in terms of simplicity, speed, throughput, and potential for robust quantitative analysis. Existing techniques that accommodate solid-phase samples incorporating individual micron-sized targets generally rely on digital microscopy and image analysis, with in...

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Automated detection of rare‐event pathogens through time‐gated luminescence scanning microscopy

Cited by 22 publications

References 26 publications

Demonstration of true‐color high‐contrast microorganism imaging for terbium bioprobes

Demonstration of true‐color high‐contrast microorganism imaging for terbium bioprobes

Cytometric investigation of rare-events featuring time-gated detection and high-speed stage scanning

High-Precision Pinpointing of Luminescent Targets in Encoder-Assisted Scanning Microscopy Allowing High-Speed Quantitative Analysis

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