Journal: Journal of Nanobiotechnology

Article Title: Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

doi: 10.1186/s12951-015-0097-1

Figure Lengend Snippet: Characterization of the designed QD-BRET nanoparticles. Nanoparticles were analyzed with transmission electron microscopy (TEM), dynamic light scattering (DLS), and agarose gel electrophoresis. The TEM image shows the core–shell of the quantum dot nanocrystals averaging 8.2 ± 1.7 (SD, of 20 measurements); while the DLS measured the diameter sizes (n = 5) of the functionalized (32 ± 1.3 nm) vs. non-functionalized (26 ± 1.3 nm) QD-BRET that were significantly different ( a , b ; P < 0.05; T test). The agarose gel shows differential migration of both nanoparticles, with functionalized aliquots (1+ , 2+ , and 5+) being heavier and migrating slowly than their non-functionalized counterparts (1−, 2−, and 5−). The gel was loaded with various concentrations of nanoparticles (1, 2, and 5 nM/well, equivalent to 3 × 10 11 , 6 × 10 11 , and 15 × 10 11 nanoparticles, respectively). A 100 base pair (bp) PCR DNA ladder (Lad.) and sperm genomic DNA (gDNA) were loaded alongside for quality control of the gel electrophoresis.

Article Snippet: The calculated average (±SD) of random measurements of QD core–shell (CdSe/ZnS) diameters through the transmission electron microscopy (TEM) images was 8.2 ± 1.7 nm, which appeared little bit higher than our previous report [ ] using a different batch of QD-BRET nanoparticles prepared by the same company (Zymera, Inc.).

Techniques: Transmission Assay, Electron Microscopy, Agarose Gel Electrophoresis, Migration, Control, Nucleic Acid Electrophoresis

Journal: Journal of Nanobiotechnology

Article Title: Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

doi: 10.1186/s12951-015-0097-1

Figure Lengend Snippet: Flow cytometry evaluations of sperm labeling and viability. Spermatozoa were labeled with 0 nM (Control), 0.1 nM (QD0.1−), and 1 nM (QD1− and QD1+) nanoparticles, followed by their incubation with FITC-PSA to the acrosome integrity or intactness. Proportions of 0%, 76 ± 4%, and 91 ± 2% spermatozoa were labeled with nanoparticles in the Control, QD0.1, QD1−, and QD1+ groups. The mean fluorescence intensities of nanoparticles-labeled spermatozoa ( A ) and proportions of spermatozoa with intact and damage acrosomes ( B ) were evaluated. Spermatozoa incubated with 0 and 10 µM calcium ionophore served as negative and positive controls, respectively. Columns (RFI, in A and QD+ intact acrosome, in B ) with different letters differ significantly (ANOVA-1; p < 0.05). Data are mean ± SEM of four independent replicates.

Article Snippet: The calculated average (±SD) of random measurements of QD core–shell (CdSe/ZnS) diameters through the transmission electron microscopy (TEM) images was 8.2 ± 1.7 nm, which appeared little bit higher than our previous report [ ] using a different batch of QD-BRET nanoparticles prepared by the same company (Zymera, Inc.).

Techniques: Flow Cytometry, Labeling, Control, Incubation, Fluorescence

Journal: Journal of Nanobiotechnology

Article Title: Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

doi: 10.1186/s12951-015-0097-1

Figure Lengend Snippet: Confocal fluorescence imaging of QD-BRET labeled spermatozoa. Spermatozoa were labeled with 0 nM (control) and 1 nM plasminogen-conjugated (QD1+) or non-conjugated (QD1−) QD-BRET nanoparticles. The control group was used for fluorescence imaging settings and was designated as the negative control group. These settings were used to capture the fluorescence emission ( upper panel ). Both fluorescence and visible light signals are overlaid in the bottom panel. Scale bar 10 µm.

Article Snippet: The calculated average (±SD) of random measurements of QD core–shell (CdSe/ZnS) diameters through the transmission electron microscopy (TEM) images was 8.2 ± 1.7 nm, which appeared little bit higher than our previous report [ ] using a different batch of QD-BRET nanoparticles prepared by the same company (Zymera, Inc.).

Techniques: Fluorescence, Imaging, Labeling, Control, Negative Control

Journal: Journal of Nanobiotechnology

Article Title: Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

doi: 10.1186/s12951-015-0097-1

Figure Lengend Snippet: Confocal fluorescence imaging of QD-BRET (QD)-labeled cumulus-oocyte complexes (COCs). COCs were labeled with 0 nM (QD0) or with 1 nM plasminogen-conjugated (QD1+) or non-conjugated (QD1−) QD-BRET nanoparticles. Non-labeled mature or immature COCs were used to set up the imaging conditions ( a , b ). Micrographs in the upper panel ( Blue frame ) show QD-BRET fluorescence signals detected in COCs labeled before (Immature; c , d ) and after (Mature; e , f , g , h ) in vitro maturation. The lower panel ( Black frame ) shows corresponding overlaid visible and fluorescence light images. Nuclei are counterstained in blue with DAPI. The white and red arrows indicate the cumulus cells and oocytes, respectively.

Article Snippet: The calculated average (±SD) of random measurements of QD core–shell (CdSe/ZnS) diameters through the transmission electron microscopy (TEM) images was 8.2 ± 1.7 nm, which appeared little bit higher than our previous report [ ] using a different batch of QD-BRET nanoparticles prepared by the same company (Zymera, Inc.).

Techniques: Fluorescence, Imaging, Labeling, In Vitro

Journal: Journal of Nanobiotechnology

Article Title: Self-illuminating quantum dots for non-invasive bioluminescence imaging of mammalian gametes

doi: 10.1186/s12951-015-0097-1

Figure Lengend Snippet: Fluorescence imaging of follicular micro-sections following QD-BRET (QD) labeling. The upper panel shows the intracellular progression of the fluorescence signals of QD1− nanoparticles during culture (Day 1, 2, and 4). Fluorescence signals were mainly detected in the mural granulosa cells, surrounding the follicular cavity, on Day 1 post microinjection. The fluorescence signal progressed into the mid-section of the follicle wall (theca internal cells) on Day 2, and then the entire follicle wall, including the theca external cells on Day 4. The bottom panel represents the corresponding overlays fluorescence and visible lights. The intra-follicular COCs also incorporated the QD1− (200×). Arrows indicate the extent of tissue layers ( GC granulosa cells, TI theca internal, TE theca external). The white arrows show detached and stained granulosa cells within the follicular antrum (FA).

Article Snippet: The calculated average (±SD) of random measurements of QD core–shell (CdSe/ZnS) diameters through the transmission electron microscopy (TEM) images was 8.2 ± 1.7 nm, which appeared little bit higher than our previous report [ ] using a different batch of QD-BRET nanoparticles prepared by the same company (Zymera, Inc.).

Techniques: Fluorescence, Imaging, Labeling, Microinjection, Staining