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Image Search Results
Journal: Fertility and sterility
Article Title: Metabolic imaging using Fluorescence Lifetime Imaging Microscopy (FLIM) accurately detects mitochondrial dysfunction in mouse oocytes
doi: 10.1016/j.fertnstert.2018.07.022
Figure Lengend Snippet: Comparative assessments of Clpp+/+ and Clpp−/− oocytes. A) Brightfield images show gross morphological features of oocytes, while NADH FLIM intensity images reflect the mitochondrial distribution. B) Obvious defects in Clpp−/− oocytes were not observed with brightfield, but fluorescence imaging revealed aberrant mitochondrial distributions in many oocytes. C) Metabolic imaging of Clpp+/+ (n=55) and Clpp−/− oocytes (n=52) detected highly significant differences in five of the eight parameters measured. Parameters are plotted in order of decreasing separation, with asterisks indicating the following p-values: (*: p<10−11) and (**: p<10−26) D) If the three most sensitive metabolic parameters (NADH long lifetime, NADH fraction engaged, and FAD long lifetime) are represented in a 3D plot, we can fit a plane that perfectly separates the two data sets. E) Clpp+/+ oocytes (n=46) and Clpp−/− oocytes (n=43) were lysed to obtain individual mtDNA copy number measurements. T-tests on mtDNA measurements showed only a marginally significant difference with a p-value of 0.042. Error bars represent standard errors.
Article Snippet: Disclosures: Tim Sanchez and Dan Needleman are co-founders and stake holders at
Techniques: Fluorescence, Imaging
Journal: Fertility and sterility
Article Title: Metabolic imaging using Fluorescence Lifetime Imaging Microscopy (FLIM) accurately detects mitochondrial dysfunction in mouse oocytes
doi: 10.1016/j.fertnstert.2018.07.022
Figure Lengend Snippet: Comparative assessments of oocytes from old (12-month) and young (3-month) mice. A,B) Neither brightfield images nor FLIM intensity images revealed obvious differences in morphology or mitochondrial distribution. 20um bar. C) Conversely, metabolic imaging measurements on the same oocytes effectively differentiated young (n=35) and old (n=29) groups, with four of the eight parameters showing significant differences. Parameters are plotted in order of decreasing separation, with asterisks indicating the following p-values: (*:p<0.02) and (**: p<10−3). P-values were not as low as with Clpp, the more severe case of metabolic dysfunction. D) If the three most sensitive metabolic parameters (NADH intensity NADH short lifetime, and NADH long lifetime) are represented in a 3D plot, we can draw a plane that effectively separates the two data sets; however, there is more overlap between the distributions than in the more extreme case of Clpp. E) mtDNA copy number measurements were taken on young (n=35) and old (n=29) oocytes, and no significant difference between the two groups was observed (p=0.14). Error bars represent standard errors.
Article Snippet: Disclosures: Tim Sanchez and Dan Needleman are co-founders and stake holders at
Techniques: Imaging
Journal: Fertility and sterility
Article Title: Metabolic imaging using Fluorescence Lifetime Imaging Microscopy (FLIM) accurately detects mitochondrial dysfunction in mouse oocytes
doi: 10.1016/j.fertnstert.2018.07.022
Figure Lengend Snippet: Evaluation of safety for varying photodoses of FLIM illumination. Top: Reactive oxygen species levels were measured via HC-DCFDA fluorescence (custom units). Significant differences between illuminated and non-illuminated embryos were not observed for any of the photodoses studied. Embryos exposed to 30mM H2O2 were measured as a positive control. Error standard error bars represent variation between individual embryo measurements. Bottom: Embryos were cultured on the microscope, and blastocyst development rates of illuminated embryos were compared to non-illuminated embryos in the same dish. Embryos cultured in a standard incubator were used as a control. FLIM illumination did not have any significant impact on blastocyst development rates. Standard error bars represent variation between experiment batches.
Article Snippet: Disclosures: Tim Sanchez and Dan Needleman are co-founders and stake holders at
Techniques: Fluorescence, Positive Control, Cell Culture, Microscopy, Control