Orca er c4742 95

VEGF and p-VEGFR2 were examined on 8 μm cryosections (on day 7 after laser photocoagulation). The cryosections were blocked with 1% bovine serum albumin for 4 h at RT, then incubated with VEGF antibody (1:50) and p-VEGFR2 antibody (1:50) at 4 °C overnight. For VEGF and p-VEGFR2 staining, antigen retrieval was obtained through heated water bath at 37 °C for 10 min. Thereafter, the slides were stained with Alexa Fluor 488-conjugated goat anti-rabbit IgG (1:200; #A27034, Thermo Fisher Scientific), Alexa Fluor 546-conjugated goat anti-mouse IgG (1:200; #A11030, Thermo Fisher Scientific), and DAPI (1:500). The photomicrographs were taken by a digital high-sensitivity camera (Hamamatsu, ORCA-ER C4742-95, Japan).

Excitation
of the Pdots samples was performed with pump pulses generated via
an optical parametric amplifier from 800 nm pulses, which were converted
to 740 nm and subsequently doubled to attain 370 nm with an OPO crystal.
Aqueous solutions of Pdots were measured using a 1 cm quartz cuvette
and an excitation power at the sample of 300 μW. Fluorescence
at a right angle to the excitation was passed through a Bruker SPEC
250IS spectrograph (ca. 200 nm observation window) and onto the streak
camera (Hamamatsu streak camera and blanking unit C5680 in combination
with a Synchroscan Unit M5675). A charge-coupled device (CCD) camera
(Hamamatsu Orca-ER C4742–95) was used in the binning mode (2
× 2 pixels) to give a 512 × 512 pixel matrix. The observed
time window in time range 2 was 700 ps, while in time range 4, it
was 2000 ps with FWHM instrument response functions of 30 or 45 ps.
For the decay curves obtained from the streak camera, the procedure
written in Matlab was employed assuming a Gaussian instrument response
function. Time-correlated single photon counting (TCSPC) measurements
were carried out in a time window of 50 ns using a pulsed diode laser
source (Edinburgh Instruments EPL470, λ_exc. = 405
nm).^{32 (link)}

Root tip squashes from germinating seedlings of T. aestivum cv. Brigand 8/2 and M₃ seedlings of lines 12A5, 12H3 and 7C12 were prepared as described in Gill et al. (1991 (link)) with minor modifications.
Fluorescence in situ hybridisation (FISH) was carried out as described in Prieto et al. (2007 (link)). The probe used was a dUTP-digoxigenin labelled product generated by amplification of high quality DNA of Ae. sharonensis accession 1644 with UTV39 primers. Labelling with dUTP-digoxigenin was carried out directly, with no nick translation being necessary due to the short length of the PCR product. Observations were made using a Nikon Eclipse E600 CCD microscope and images captured with a Hamamatsu ORCA-ER C4742-95 digital camera. Images were merged using ImageJ software.