Natural and artificial pollination of white-fleshed pitaya.

To produce pitaya (Hylocereus spp.), it is essential that pollination occur, either artificially or by pollinating agents. This study evaluated the viability of self-pollination, cross-pollination, and natural pollination, as well as pollen viability, stigma receptivity and ideal pollination window in pitaya flowers (Hylocereus spp.). An experiment was conducted with a randomized block design consisting of four treatments: T1: manual self-pollination; T2: nocturnal open pollination; T3: diurnal open pollination; and T4: manual cross-pollination - red-fleshed pitaya [ Hylocereus polyrhizus] pollen placed on white-fleshed pitaya [ Hylocereus undatus] stigma. The experiment had four replicates, with two plants per replicate (two flowers per plant), totaling 16 flowers per treatment. The analyzed variables were fruit weight, length, diameter, titratable acidity, pH, soluble solids, and SS/TA ratio; the germination percentage of pollen grains; and the receptivity of stigmas. Artificial pollination (self- and cross-pollination) of white-fleshed pitaya (Hylocereus undatus) is viable, resulting in larger fruits than natural pollination, with cross-pollination being the most recommended. Throughout the floral opening period, the pitaya flower (Hylocereus spp.) has receptive stigma capable of receiving pollen. The flowers exhibit the highest pollen germination rate at 7 p.m, which is the ideal pollination window for pitaya.

Imaging of photoacoustic-mediated permeabilization of giant unilamellar vesicles (GUVs).

Target delivery of large foreign materials to cells requires transient permeabilization of the cell membrane without toxicity. Giant unilamellar vesicles (GUVs) mimic the phospholipid bilayer of the cell membrane and are also useful drug delivery vehicles. Controlled increase of the permeability of GUVs is a delicate balance between sufficient perturbation for the delivery of the GUV contents and damage to the vesicles. Here we show that photoacoustic waves can promote the release of FITC-dextran or GFP from GUVs without damage. Real-time interferometric imaging offers the first movies of photoacoustic wave propagation and interaction with GUVs. The photoacoustic waves are seen as mostly compressive half-cycle pulses with peak pressures of ~ 1 MPa and spatial extent FWHM ~ 36 µm. At a repetition rate of 10 Hz, they enable the release of 25% of the FITC-dextran content of GUVs in 15 min. Such photoacoustic waves may enable non-invasive targeted release of GUVs and cell transfection over large volumes of tissues in just a few minutes.

Photoacoustic generation of intense and broadband ultrasound pulses with functionalized carbon nanotubes.

Carbon nanotubes (CNT) functionalized with siloxane groups were dissolved in polystyrene/tetrahydrofuran to produce thin films that generate broadband and intense ultrasound pulses when excited by pulsed lasers. These films absorb >99% of light in the visible and near-infrared and show no signs of fatigue after thousands of laser pulses. Picosecond laser pulses with fluences of 50 mJ cm-2 generate photoacoustic waves with exceptionally wide bandwidths (170 MHz at -6 dB) and peak pressures >1 MPa several millimeters away from the source. The ability to generate such broadband ultrasound pulses is assigned to the ultrafast dissipation of heat by CNT-siloxanes, and to the formation of very thin photoacoustic sources thanks to the high speed of sound of polystyrene. The wide bandwidths achieved allow for axial resolutions of 8 µm at depths less than 1 mm, similar to the resolution of histology but based on real-time non-invasive methods.

Photoacoustic transfection of DNA encoding GFP.

Photoacoustic transfection consists in the use of photoacoustic waves, generated in the thermoelastic expansion of a confined material absorbing a short pulse of a laser, to produce temporary mechanical deformations of the cell membrane and facilitate the delivery of plasmid DNA into cells. We show that high stress gradients, produced when picosecond laser pulses with a fluence of 100 mJ/cm2 are absorbed by piezophotonic materials, enable transfection of a plasmid DNA encoding Green Fluorescent Protein (gWizGFP, 3.74 MDa) in COS-7 monkey fibroblast cells with an efficiency of 5% at 20 °C, in 10 minutes. We did not observe significant cytotoxicity under these conditions. Photoacoustic transfection is scalable, affordable, enables nuclear localization and the dosage is easily controlled by the laser parameters.

Phthalocyanine Labels for Near-Infrared Fluorescence Imaging of Solid Tumors.

Diamagnetic metal complexes of phthalocyanines with n-butoxyl groups in all the α-benzo positions of the macrocycle skeleton, MPc(OBu)8, have strong near-infrared absorptions and intense fluorescences that are Stokes shifted by more than 15 nm. Interestingly, the silicon complex 6 is also remarkably photostable and nontoxic. The use of 6 in the fluorescence imaging of BALB/c mice bearing a 4T1-luc2 tumor in the mammary fat pad unambiguously revealed the presence of the tumor when it was only 1 mm in diameter and was not visible with the naked eye. Compound 6 has an intrinsic ability to accumulate in the tumor, adequate spectroscopic properties, and excellent stability to function as a NIR fluorescent label in the early detection of tumors.