An electronically cooled camera from Princeton Instruments (PIXIS 1024) was used to follow luminescence using the Lightfield program. Camera runs were completed at 25°C, with cultures grown in black, flat-bottomed 96-well plates from Eppendorf. Liquid suspensions of conidia (O.D. of 0.5 at 600nm) were plated onto QA medium (0.03% glucose, 0.05% arginine, 0.001 M QA, pH 5.75) containing 25mM luciferin and covered with a Breathe-Easy strip (USAScientific). Inoculated plates were then subjected to 48 h of 12:12 dark:light cycle conditions at 25°C:28°C before starting the camera trial in constant-dark and 25°C conditions. Signals were accumulated for 15 min every hour. A custom ImageJ Macro called “Toolset Image Analysis Larrondo’s Lab v. 1.0” was used to process the images (Larrondo et al., 2012 (link)) within FIJI v.2.0.0 (Schindelin et al., 2012 (link)). Raw data arising from each time series was smoothed using a 3-point moving average, and then processed using custom-written software to detrend (by removing the exponential decay signal), rescale, and normalize amplitudes.
Pixis 1024
Manufactured by Teledyne
The Pixis 1024 is a digital CCD camera designed for laboratory applications. It features a 1024 x 1024 pixel sensor with a pixel size of 13 x 13 microns. The camera provides 16-bit image depth and a maximum readout rate of 2 MHz.
Automatically generated - may contain errors
7 protocols using pixis 1024
1
Bioluminescence Assay for Fungal Circadian Rhythms
2
Single-Cell RNA FISH Imaging Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
RNA FISH was carried out largely as reported previously15 (link). Cells were dissociated by trypsinization, washed in PBS, fixed in 4% paraformaldehyde at room temperature and permeabilized and stored in 70% ethanol at 4 °C. All washes and hybridizations were carried out in suspension. Wash buffers included 0.1% Triton X-100 to minimize losses to sticking on the walls. Samples were mounted between coverglasses in the glucose-oxidase-based 2 × SSC anti-fade buffer described previously15 (link). We imaged using a 100 × 1.4NA oil-immersion objective, a Nikon Ti-E wide field microscope, and a deep-depletion CCD camera (Pixis 1024, Princeton Instruments).
3
Single-molecule FISH for Caenorhabditis elegans
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
smFISH experiments were performed as described elsewhere (Raj et al. 2008 (link)). Fixed mixed stage animals were incubated overnight at 30° with previously designed egl-1 probes (Johnsen and Horvitz 2016 (link)). Animals were subsequently mounted for imaging after two washes lasting 30 min each. Image acquisition was performed on a Nikon TE-2000 inverted microscope with a 100x objective (Nikon, NA 1.4). A Pixis 1024 camera (Princeton Instruments) controlled by MetaMorph software (Molecular Devices) was used to detect smFISH signal and acquire images with an exposure time of 2 sec. Images were processed and prepared for publication using ImageJ software (NIH).
4
Widefield Microscopy of Cell Samples
5
Optical Characterization of Silver Nanowire Antennas
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Light-scattering measurements on the silver antennas protruding from the junctions were carried out using a confocal optical microscope coupled to a true-colour CCD (Nikon) and a spectrometre (Spectra Physics) with a cooled CCD (Princeton Instruments, Pixis 1,024). The silver nanowire devices were illuminated in the bright-field objective (Nikon NA0.9, × 100) with white light from a halogen lamp. The light from the silver nanowire antenna was confocally selected and sent to the spectrometre to acquire quantitative spectral information. The spectral data were normalized by subtracting the spectrum backscattered from the air–glass interface from the spectrum of the antenna, then dividing by the lamp spectrum. The spatial resolution of the confocal collection scheme was ∼1 μm. The estimated resistance of the metallic leads and the nanowire of the cross-point junction where calculated using COMSOL Multiphysics using the electric currents module for a device of the same dimensions as the device measured in the optical and STEM EELS experiments.
6
Single-Molecule Fluorescence In Situ Hybridization
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Probe design and smFISH experiments were performed essentially as described elsewhere [19 (link)] with 10% formamide in the hybridization buffer. Samples were incubated overnight at 30˚C and washed for 30 min twice. Imaging was performed using an inverted microscope with a 100x oil objective (Nikon, NA 1.4). smFISH signal was detected using a Pixis 1024 (Princeton Instruments) with exposure times of 2 sec. For msp FISH the high fluorescence intensity required exposure times of 500 msec. and a scMos Orca Flash 4.0 camera (Hamamatsu) was used for fluorescence detection.
7
Nanodisk Dimer Characterization via Confocal Microscopy
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
We use a confocal microscope (Nikon C2 Plus confocal microscope) with a dark-field module. The light from a tungsten-halogen lamp illuminates the sample through a dark-field condenser and a 100× objective lens (Nikon LU Plan Fluor 100× BD). The scattered light from the gold nanodisk dimer is collected by the same objective lens and the signal is transferred by an optical fiber connected to a spectrometer (SpectraPro, Acton 2300i). We use a Si array detector (Princeton Instruments, Pixis 1024) for the visible range measurements and an InGaAs array detector (Princeton Instruments, 7498-0001) for the near-infrared range measurements.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!