High intensity solid‐state UV source for time‐gated luminescence microscopy

Connally, Russell; Jin, Dayong; Piper, James A.

doi:10.1002/cyto.a.20326

Cited by 46 publications

(71 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Output power and temporal parameters of LEDs can be controlled easily by simple electronics in contrast to flash lamp electronics. The technological advancement of LEDs has already resulted in the development of the lanthanide chelates based time-resolved systems using LED excitation at 365 nm [9]. A 100 mW single LED at 365 nm with pulse duration of 101 μs and a gate delay of 5 μs was employed for excitation in time-gated luminescence microscopy [9].…”

Section: Introductionmentioning

confidence: 99%

“…The technological advancement of LEDs has already resulted in the development of the lanthanide chelates based time-resolved systems using LED excitation at 365 nm [9]. A 100 mW single LED at 365 nm with pulse duration of 101 μs and a gate delay of 5 μs was employed for excitation in time-gated luminescence microscopy [9]. LEDs at 365 nm were also used in a time-resolved luminescence microscope for pulsed epi-illumination [10].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays

Rodenko¹,

Fodgaard²,

Tidemand-Lichtenberg³

et al. 2016

Opt. Express

View full text Add to dashboard Cite

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

340 nm pulsed UV LED system for europium-based time-resolved fluorescence detection of immunoassays

Rodenko¹,

Fodgaard²,

Tidemand-Lichtenberg³

et al. 2016

Opt. Express

View full text Add to dashboard Cite

“…The UV optics match the peak absorption (300-330 nm) for the most biocompatible terbium complexes (9,18,21). A 0.5-mW, 315 nm LED source (UVTOP310 with a hemispherical lens window, Sensor Electronic Technology, Columbia, SC, USA, http://www.s-et.com) replaced the previously reported highpower, 365 nm (UV) LED (1,31); since the typical objective lens and cover slip glass cut-off transmission is below 330 nm, a MicroSpot Focusing UV objective 403, 240-360 nm, and NA 5 0.50 (Thorlabs, LMU-40X-UVB, http://www.thorlabs. com/) was used for epi-illumination.…”

Section: Methodsmentioning

confidence: 99%

“…The sample was then washed three times with the mAb buffer. Subsequently, 20 lL of the diluted LanthaScreen 1 Tb-Streptavidin conjugate solution (50 lg/mL) was added and allowed to react for another 2 hr. This was followed by one wash with 400 lL of the mAb buffer to prepare the Cryptosporidium parvum oocysts in 100 lL suspension for microscopy imaging.…”

Section: Cryptosporidium Labelingmentioning

confidence: 99%

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

Jin

2011

Cytometry Pt A

Self Cite

View full text Add to dashboard Cite

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...

show abstract

“…Also in this issue, a multicontinent collaboration is reported by Yiqing Lu et al describing an approach for the automated detection of rare-event pathogens through time-gated luminescence scanning microscopy. It has been recognized that luminescence microscopes operating in the time-domain can improve the detection of signals from long fluorescence lifetimes (tau > 100 microseconds) in a background of short-lived autofluorescence lifetime noise frequently observed in micro-organisms (10). The team addressed the need to improve methods for the rapid detection of infectious disease, in particular monitoring waterborne pathogenic microorganisms in recreational waters (typically, Giardia lamblia cysts and Cryptosporidium parvum oocysts).…”

mentioning

confidence: 99%