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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.
  1. 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.