Steady state photoluminescence (PL) spectra of the films were recorded in a fluorimeter (Edinburgh Instruments, FLS980). Transient PL decay curves were recorded on the same fluorimeter by time-correlated-single-photon-counting (TCSPC). Excitation was by a 379 nm laser diode (Pico Quant, LDH-D-C-375) operating at a repetition rate of 300 kHz. The measured PL decay curves were then fitted by a two exponential decay model considering the instruments response function (IRF) and an averaged emission lifetime (<τ>) of the films was estimated using the following equation; , where τ1 and τ2 are the emission lifetime of each component and γ1 and γ1 are the contribution of the emission from each component to the total emission (i.e., , where A1 and A2 are the pre-exponential-factors of each component). PL quantum yield of the films were measured on unencapsulated films using an integrating sphere based measurement system (Hamamatsu, C9920-02) under nitrogen flow.
Ldh d c 375
Manufactured by PicoQuant
Sourced in Germany
The LDH-D-C-375 is a pulsed diode laser from PicoQuant designed for use in time-resolved fluorescence spectroscopy and imaging applications. The laser operates at a wavelength of 375 nm and provides picosecond pulse widths. The output power and repetition rate can be adjusted to suit the specific requirements of the experiment.
Automatically generated - may contain errors
Lab products found in correlation
Fluofit, by PicoQuant (2 mentions)
Timeharp platine, by PicoQuant (2 mentions)
Monochromator spectrapro hrs 300, by Teledyne (2 mentions)
Fls980, by Edinburgh Instruments (1 mentions)
C9920 02, by Hamamatsu Photonics (1 mentions)
Ixon ultra 897, by Oxford Instruments (1 mentions)
Ti2 a inverted microscope, by Nikon (1 mentions)
Fesh0450, by Thorlabs (1 mentions)
Felh0500, by Thorlabs (1 mentions)
R3809u 50 microchannel plate photomultiplier, by Hamamatsu Photonics (1 mentions)
Fluorolog 3 spectrofluorometer, by Horiba (1 mentions)
Fs5 spectrofluorometer, by Edinburgh Instruments (1 mentions)
Pda100a, by Thorlabs (1 mentions)
Pma hybrid 40, by PicoQuant (1 mentions)
Symphotime 64, by PicoQuant (1 mentions)
Microtime 200 microscope, by PicoQuant (1 mentions)
6 protocols using ldh d c 375
1
Photoluminescence Characterization of Thin Films
2
Fluorescence Microscopy Experimental Setup
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Fluorescence
microscopy experiments were performed using a Nikon Ti2-A inverted
microscope. The samples are placed on the microscope’s table,
which was applied with a custom-made box to allow measurements under
inert conditions. The excitation light was from a 373 nm collimated
free-beam laser diode (LDH-D-C-375, PicoQuant), passing a clean-up
filter (370/36 BrightLine HC, Semrock) and a lambda fourth plate (355
nm, Edmund Optics). The beam was expanded using a 10× UV beam
expander (BE10-UVB, Thorlabs, Inc.) and then focused on the back-focal
plane of the objective to enable far-field microscopy. It entered
the microscope through the backside port and was mirrored to the sample
stage via a dichroic mirror (zt 375 RDC, Chroma). Emitted light from
the sample was collected by the objective and passed the dichroic
mirror to be led to a side port of the microscope. Here, it was spectrally
separated into two parts using color filters (FESH0450 and FELH0500,
Thorlabs) and a dichroic mirror (zt 514 RDC, Chroma) mounted on an
Optosplit II (Acal BFi Germany GmbH). The two resulting images represented
the wavelength regimes. The image detection was done using a back-illuminated
CCD camera (iXon Ultra 897, Andor). Time-resolved measurements were
realized by taking a series of images and subsequent post-procession
of the data with a self-written evaluation script.
microscopy experiments were performed using a Nikon Ti2-A inverted
microscope. The samples are placed on the microscope’s table,
which was applied with a custom-made box to allow measurements under
inert conditions. The excitation light was from a 373 nm collimated
free-beam laser diode (LDH-D-C-375, PicoQuant), passing a clean-up
filter (370/36 BrightLine HC, Semrock) and a lambda fourth plate (355
nm, Edmund Optics). The beam was expanded using a 10× UV beam
expander (BE10-UVB, Thorlabs, Inc.) and then focused on the back-focal
plane of the objective to enable far-field microscopy. It entered
the microscope through the backside port and was mirrored to the sample
stage via a dichroic mirror (zt 375 RDC, Chroma). Emitted light from
the sample was collected by the objective and passed the dichroic
mirror to be led to a side port of the microscope. Here, it was spectrally
separated into two parts using color filters (FESH0450 and FELH0500,
Thorlabs) and a dichroic mirror (zt 514 RDC, Chroma) mounted on an
Optosplit II (Acal BFi Germany GmbH). The two resulting images represented
the wavelength regimes. The image detection was done using a back-illuminated
CCD camera (iXon Ultra 897, Andor). Time-resolved measurements were
realized by taking a series of images and subsequent post-procession
of the data with a self-written evaluation script.
3
Steady-state and Time-resolved Fluorescence Analysis
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Steady-state excitation and emission spectra were acquired using a Fluorolog-3 spectrofluorometer (model FL3-11; Jobin Yvon, Edison, NJ) and FS-5 spectrofluorometer (Edinburgh Instruments, UK) equipped with a xenon arc lamp. The temperature in the cuvette holders was maintained using a water-circulating bath. The steady-state spectra were recorded in steps of 1 nm (bandwidths of 1.2 nm were chosen for both the excitation and emission monochromators) in triplicates and averaged.
Fluorescence decays were recorded on a 5000 U single-photon counting setup (IBH, Glasgow, UK) using a laser excitation source (peak wavelengths of 375 nm (LDH-DC-375, Picoquant, Germany) and 532 nm (PicoTa, Toptica Photonics AG, Germany), for Pro12A and NR-probes, respectively, at a 5 MHz repetition rate) and a cooled R3809U-50 microchannel plate photomultiplier (Hamamatsu, Japan). A 399 nm or 550 nm cut-off filter was used to eliminate scattered light. The signal was kept below 1% of the repetition rate of the light source. Data were collected until the peak value reached 5000 counts. Measurements were performed at 23°C and 37°C.
Fluorescence decays were recorded on a 5000 U single-photon counting setup (IBH, Glasgow, UK) using a laser excitation source (peak wavelengths of 375 nm (LDH-DC-375, Picoquant, Germany) and 532 nm (PicoTa, Toptica Photonics AG, Germany), for Pro12A and NR-probes, respectively, at a 5 MHz repetition rate) and a cooled R3809U-50 microchannel plate photomultiplier (Hamamatsu, Japan). A 399 nm or 550 nm cut-off filter was used to eliminate scattered light. The signal was kept below 1% of the repetition rate of the light source. Data were collected until the peak value reached 5000 counts. Measurements were performed at 23°C and 37°C.
4
Fluorescence Lifetime Measurement Methods
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Lifetimes in the ms regime were determined using a 340 nm M340L4 LED or a 365 nm LED M365L2 (Thorlabs), a TGP3122 pulse generator (AIM‐TTI Instruments), and a silicon photodetector PDA100A (Thorlabs). For the ns and µs regime, a time correlated single photon counting (TCSPC) setup was used, containing a 375 nm laser LDHDC375, a PMA Hybrid Detector PMA Hybrid 40, a TimeHarp platine (all PicoQuant), and a Monochromator SpectraPro HRS‐300 (Princeton Instruments). ns‐Lifetimes were evaluated using reconvolution algorithms of FluoFit (PicoQuant).
5
TCSPC Setup for Perovskite Film Analysis
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A TCSPC setup is composed of a 375 nm laser diode head (Pico Quant LDHDC375), a PMA Hybrid detector (PMA Hybrid 40), a TimeHarp platine (all PicoQuant), and a Monochromator SpectraPro HRS-300 (Princeton Instruments). Perovskite films on quartz were excited with the 375 nm laser diode and then the emission was collected by using the PMA hybrid detector. The pulse width is ≈44 ps, the power is ≈3 mW, and the spot size is ≈1 mm2, so the excitation fluence is ≈0.132 J m−2. The lifetimes were evaluated using reconvolution algorithms of FluoFit (PicoQuant).
6
Fluorescence Lifetime Imaging of ACB10 in MDA-MB-231 Cells
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MDA-MB-231 cell lines were grown in Dulbecco’s modified Eagle medium (DMEM). Cells were seeded onto 20 mm square glass cover slides into 6-well plates and cultured (2.5 × 105 cells per plate) at 37 °C in a 5% CO2 humidified atmosphere with their respective medium. The cells were incubated with 5 μM of ACB10 in DMEM medium without phenol red for 1, 4, 24, and 48 h. After incubation, the cells were washed three times with PBS.
Fluorescence lifetime images were recorded with a MicroTime 200 microscope (PicoQuant) equipped with a TCSPC card and two TAU-SPAD-100 avalanche photodiode detectors. A 375 nm pulsed diode laser (LDH-D-C-375, PicoQuant) was used as an excitation source, at a 10 MHz repetition rate and a power of ~0.7 µW. The emission was recorded with a long-pass filter (−519/19 LP). The regions of 80 × 80 µm were scanned with 156 nm/pixel spatial resolution and 2 ms of dwell time. FLIM images were processed using SymphoTime64 software (PicoQuant, Berlin, Germany). The lifetime distribution histograms were obtained from FLIM images and were fitted to the Gaussian curve. The FLIM images were smoothed over 200 nm for clarity of presentation. The emission spectra and the histograms were averaged over 3 independent measurements.
Fluorescence lifetime images were recorded with a MicroTime 200 microscope (PicoQuant) equipped with a TCSPC card and two TAU-SPAD-100 avalanche photodiode detectors. A 375 nm pulsed diode laser (LDH-D-C-375, PicoQuant) was used as an excitation source, at a 10 MHz repetition rate and a power of ~0.7 µW. The emission was recorded with a long-pass filter (−519/19 LP). The regions of 80 × 80 µm were scanned with 156 nm/pixel spatial resolution and 2 ms of dwell time. FLIM images were processed using SymphoTime64 software (PicoQuant, Berlin, Germany). The lifetime distribution histograms were obtained from FLIM images and were fitted to the Gaussian curve. The FLIM images were smoothed over 200 nm for clarity of presentation. The emission spectra and the histograms were averaged over 3 independent measurements.
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