Non invasive in vivo investigation of hepatobiliary structure and function in STII medaka (Oryzias latipes): methodology and applications

Hardman, Ron C; Kullman, Seth W; Hinton, David E

doi:10.1186/1476-5926-7-7

Table 1 Fluorescent probe specifications

From: Non invasive in vivo investigation of hepatobiliary structure and function in STII medaka (Oryzias latipes): methodology and applications

Fluorophore	Soluble	Ex	Em	Initial/Peak Assimilation Time (min)	Exposure Concentration
7-benzyloxyresorufin	DMSO	560	590	30/45	50 μM
In vivo CYP3A activity. Uptake via gill, in vivo metabolism in gill and gut, good for investigating blood to bile transport, IHBPs, EHBPs and intestinal lumen.
7-Ethoxyresorufin	DMSO			30/45	10–50 μM
In vivo imaging of CYP1A activity. Uptake via gill, in vivo metabolism in gill and liver. CYP 1A2, 2E Substrate
Acridine Orange	H₂O	500	526		1–5 μM
In vivo labeling of DNA, RNA. Good for apoptosis, interacts with DNA and RNA by intercalation or electrostatic attractions.
BODIPY 505/515: 4,4-difluoro-1,3,5,7-tetramethyl-4-bora-3a,4a-diaza-s-indacene	DMSO	502	510	15/20	5 μM-100 nM
Uptake via gill, active transport through hepatobiliary system, concentrative blood to bile transport, and secretion from gall bladder into gut lumen. Good for elucidation of gill, IHBP, EHBP, intestinal lumen. Non-Polar, Lipophilic.
BODIPY FL C5-ceramide: N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)sphingosine	DMSO	358	461	20/45	5 μM–100 nM
Putative passive transport in vivo. Uptake via gill, transport through cardiovascular and hepatobiliary systems and secretion from gall bladder into gut lumen.
Bodipy Verapamil	DMSO	504	511	20/60
In vivo Bodipy verapamil localized to hepatocytic cytosol in discrete vesicles. Transport to bile was not observed in the time frame assayed, which was 90 minutes.
BODIPY ^® 493/503: 4,4-difluoro-1,3,5,7,8-pentamethyl-4-bora-3a,4a-diaza-s-indacene (BODIPY ^® 493/503)	DMSO	493	504	15/20	10 μM
Uptake via gill, active transport through hepatobiliary system, concentrative blood to bile transport, and secretion from gall bladder into gut lumen. Good for elucidation of gill, IHBPs, EHBPs, intestinal lumen. Lipophilic, amphiphilic.
BODIPY ^® 581/591 C5-HPC (Phosphocholine) PC: 2-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-pentanoyl)-1-hexadecanoyl-sn-glycero-3-phosphocholine	DMSO	582	593	15/20	30 nM
OIn vivo labeling of intrahepatic and extrahepatic biliary system. Uptake via gill. Hepatobiliary transport to gut lumen. Diffuse fluorescence in hepatocyte cytosol.
CellTrace™ Oregon Green ^® *488 carboxylic acid diacetate, succinimidyl ester (carboxy-DFFDA, SE) cell permeant* mixed isomers*	DMSO	<300	none	15/20	10–100 μM
In vivo labeling of hepatic nuclei, potential for detection of apoptosis
DAPI: 4',6-diamidino-2-phenylindole, dihydrochloride	H2O, DMF	358	461	15/60	0.3 – 3.0 μM
In vivo nuclear labeling of virtually all cell types associated with gill, gut, liver, and cardiovascular system.
DAPI Diacetate: 4',6-diamidino-2-phenylindole, diacetate	H2O, MeOH	358	461	15/60	0.3 – 3.0 μM
DAPI diacetate is water-soluble form. In vivo nuclear labeling.
Fluorescein-5-isothiocyanate (FITC 'Isomer I')	DMSO	494	519	10/30	1 μM – 50 nM
Excellent in vivo probe for elucidating biliary system. In vivo labeling of intrahepatic and extrahepatic biliary system. Hepatobiliary transport to intestinal lumen. Uptake via gill.
MitoTracker ^® Green FM	DMSO	490	516	20/30	25–200 nM
In vivo labeling of hepatocytes.
SYTO ^® 16 green fluorescent nucleic acid stain	DMSO	488	518	15/30	10 nM-1 μM
In vivo nuclear labeling of epithelia and endothelia, putative in vivo probe for apoptosis.
SYTO ^® 27 green fluorescent nucleic acid stain	DMSO	495	537	15/50	10 nM-1 μM
Apoptosis.
SYTOX ^® Orange nucleic acid stain	DMSO	547	570	15/50	0.1–5 μM
Apoptosis
YO-PRO ^® -1 iodide (491/509)	DMSO		509	15/60	1 μM
Apoptosis
YOYO-1 Iodide (491/509)	DMSO	491	509	15/50	2 – 5 nM
In vivo accumulated in interstitial spaces, some labeling of vasculature. Typically used for assays for cell enumeration, cell proliferation and cell cycle.

Eighteen fluorescent probes were selected for in vivo application based on their molecular weight, utility in elucidating desired structure/function, biocompatibility, and in vivo transport properties. The fluorescent probe, a description of application(s), the molecular weight (M.W.), solvent used for stock preparation (solubility), fluorescence excitation and emission wavelengths (Ex/Em), initial and peak fluorescence times (Initial/Peak Fluorescence (min)), and effective aqueous exposure concentrations are given.

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Comparative Hepatology