Journal: Scientific Reports

Article Title: Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging

doi: 10.1038/s41598-017-07820-6

Figure Lengend Snippet: Schematic of the structure of Pink Flamindo. ( a ) Drawings for mApple and resultant Pink Flamindo. Asterisks indicate mutations. ( b ) 3D representation of Pink Flamindo bound (right) and unbound (left) to cAMP. Images were created using structural graphics from mCherry (PDB_4ZIO), which has the same origin of gene as mApple and Epac1 (cAMP-unbound: PDB_2BYV, cAMP-bound: PDB_4MGK).

Article Snippet: The adeno-associated virus (AAV) pAAV-hGFAP-Pink Flamindo and pAAV-hGFAP-Flamindo2 constructs were synthesised by replacing the promoter and coding region of the pAAV-hSyn-EGFP vector (Addgene plasmid #50465, a gift from Bryan Roth) with the human GFAP promoter and Pink Flamindo or Flamindo2 sequence; the latter was inserted between the Age I and Not I sites.

Techniques:

Journal: Scientific Reports

Article Title: Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging

doi: 10.1038/s41598-017-07820-6

Figure Lengend Snippet: Spectroscopic characterisation of Pink Flamindo. ( a ) Excitation and emission spectra of purified Pink Flamindo protein in the presence (solid line) or absence (dashed line) of 100 μM cAMP. Each fluorescent intensity (FI) was normalised to the peak of FI in the absence of cAMP. ( b ) Absorbance spectra of 30 μM Pink Flamindo in the presence (solid line) or absence (dashed line) of 100 μM cAMP. ( c ) Dose-response curve of Pink Flamindo for cAMP (●, red line) and cGMP (■, gray line). K d values calculated using the Hill equation were 7.2 μM for cAMP and 94 μM for cGMP. The peak of FI in the absence of cAMP or cGMP was normalised to 0, and the peak of FI in the presence of 1 mM cAMP was normalised to 1. The data represent the means ± standard deviation (n = 3). ( d ) Titration curves of Pink Flamindo against pH in the presence (●, solid line) or absence (○, dashed line) of 100 μM cAMP. The peak FI for each pH was normalised to the peak of FI in the presence of cAMP at pH 9.0. The data represent the means ± standard deviation (n = 3).

Article Snippet: The adeno-associated virus (AAV) pAAV-hGFAP-Pink Flamindo and pAAV-hGFAP-Flamindo2 constructs were synthesised by replacing the promoter and coding region of the pAAV-hSyn-EGFP vector (Addgene plasmid #50465, a gift from Bryan Roth) with the human GFAP promoter and Pink Flamindo or Flamindo2 sequence; the latter was inserted between the Age I and Not I sites.

Techniques: Purification, Standard Deviation, Titration

Journal: Scientific Reports

Article Title: Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging

doi: 10.1038/s41598-017-07820-6

Figure Lengend Snippet: Live cell imaging using Pink Flamindo with photoactivated adenylyl cyclase, and dual-colour imaging with G-GECO. ( a ) Sequential images and time course of fluorescence intensity of Pink Flamindo expressed in HeLa cells after consecutive exposure to 100 μM Fsk, 200 μM IBMX and 100 μM DDA. Scale bar represents 20 μm. The data represent the means ± standard deviation (n = 30 cells from 5 experiments). ( b ) Sequential images and time course of fluorescence intensity of MIN6 m9 cells expressing Pink Flamindo during the application of 25 mM glucose. Scale bar represents 20 μm. The data represent the means ± standard deviation (n = 15 cells from 4 experiments). ( c ) Sequential images and time course of fluorescence intensity of HeLa cells co-overexpressing Pink Flamindo and photoactivated adenylyl cyclase (bPAC) upon blue light laser excitation at 1.8 μW. Scale bar represents 20 μm. The data represent the mean ± standard deviation (n = 23 cells from 4 experiments). ( d ) Sequential images of MIN6 m9 cells co-overexpressing Pink Flamindo and G-GECO after consecutive exposure to 200 μM tolbutamide. Scale bar represents 20 μm. ( e ) Time course of fluorescence intensity of MIN6 m9 cells co-overexpressing Pink Flamindo and G-GECO. The data represent means ± standard deviation (n = 18 cells from 4 experiments).

Article Snippet: The adeno-associated virus (AAV) pAAV-hGFAP-Pink Flamindo and pAAV-hGFAP-Flamindo2 constructs were synthesised by replacing the promoter and coding region of the pAAV-hSyn-EGFP vector (Addgene plasmid #50465, a gift from Bryan Roth) with the human GFAP promoter and Pink Flamindo or Flamindo2 sequence; the latter was inserted between the Age I and Not I sites.

Techniques: Live Cell Imaging, Imaging, Fluorescence, Standard Deviation, Expressing

Journal: Scientific Reports

Article Title: Red fluorescent protein-based cAMP indicator applicable to optogenetics and in vivo imaging

doi: 10.1038/s41598-017-07820-6

Figure Lengend Snippet: In vivo imaging of cerebral cortical astrocytes using Pink Flamindo. ( a ) Image showing the experimental setup of a mouse under anaesthesia, positioned under a two-photon microscope. ( b ) Schematic of the cranial window for in vivo two-photon imaging. ( c ) Sequential images of cortical astrocytes expressing Pink Flamindo during exposure to 50 μM Fsk and 500 μM IBMX. Scale bar represents 100 μm. ( d ) Enlarged image of c (24 min) to indicate the regions of interest. Analysed cells are marked in red. Yellow dashed line indicates the coverslip border (see b ). Scale bar represents 50 μm. ( e ) Population mean trace of fluorescence intensity for Pink Flamindo from the images shown in c . The topical application of Fsk, IBMX and compound washout (indicated by black horizontal bars) was repeated twice. The data represent the means ± standard deviation (n = 9 cells).

Article Snippet: The adeno-associated virus (AAV) pAAV-hGFAP-Pink Flamindo and pAAV-hGFAP-Flamindo2 constructs were synthesised by replacing the promoter and coding region of the pAAV-hSyn-EGFP vector (Addgene plasmid #50465, a gift from Bryan Roth) with the human GFAP promoter and Pink Flamindo or Flamindo2 sequence; the latter was inserted between the Age I and Not I sites.

Techniques: In Vivo Imaging, Microscopy, In Vivo, Imaging, Expressing, Fluorescence, Standard Deviation

Journal: Integrative Medicine: A Clinician's Journal

Article Title: A Study on the Bioavailability of a Proprietary, Sustained-release Formulation of Astaxanthin

doi:

Figure Lengend Snippet: Comparison of the Dissolution Profiles of Astaxanthin-SR and Astaxanthin Oil

Article Snippet: The standard astaxanthin from Chromadex was used for the quantification of plasma astaxanthin.

Techniques:

Journal: Integrative Medicine: A Clinician's Journal

Article Title: A Study on the Bioavailability of a Proprietary, Sustained-release Formulation of Astaxanthin

doi:

Figure Lengend Snippet: Comparison of the Uptake of Astaxanthin From Astaxanthin-SR and Astaxanthin Oil After a Single Oral Dose

Article Snippet: The standard astaxanthin from Chromadex was used for the quantification of plasma astaxanthin.

Techniques:

Journal: Integrative Medicine: A Clinician's Journal

Article Title: A Study on the Bioavailability of a Proprietary, Sustained-release Formulation of Astaxanthin

doi:

Figure Lengend Snippet: Comparison of the Uptake of Astaxanthin From Astaxanthin-SR and Astaxanthin Oil by Participants

Article Snippet: The standard astaxanthin from Chromadex was used for the quantification of plasma astaxanthin.

Techniques: