Dear Melike,

According to the recent articles that I have read, the best way is by lyophilization:

The following text describes the procedure implemented in one of recent papers that I have read (attached):

1-In Vivo Imaging of a New Indocyanine Green Micelle

Formulation in an Animal Model of Laser-Induced

Choroidal Neovascularization

Johanna Meyer,1 Alexander Cunea,1 Dagmar Sonntag-Bensch,1 Pia Welker,2 Kai Licha,2

Frank G. Holz,1 and Steffen Schmitz-Valckenberg

ICG Dyes

Two different ICG probes were investigated herein: the

conventional, commercially available ICG dye (Pulsion, Feldkirchen,

Germany) and a newly developed encapsulated ICG

dye formulation. For the latter, ICG (Cardiogreen Standard

Fluka; Sigma-Aldrich, Hamburg, Germany) was formulated with

the nonionic solubilizer and emulsifying agent Kolliphor HS 15

(BASF, Ludwigshafen, Germany), also known as 2-hydroxyethyl

12-hydroxyoctadecanoate, to create ICG/HS 15 (mivenion

GmbH, Berlin, Germany; Figs. 1A, 1B). Kolliphor HS 15 is

amphiphilic, soluble in water, and exhibits a low toxicity and

viscosity in rats and dogs.35 One ICG molecule was encapsulated

with 2000 molecules Kolliphor HS 15 to create a stable

and spherical micelle (Fig. 1C). The micelles exhibit an average

diameter of 13 nm.36 The absorption maximum is at 798 nm,

the emission maximum is at 825 nm (in aqua dest).

The ICG/HS 15 was prepared under sterile conditions and

carried over in phosphate-buffered saline solution (PBS). After

encapsulating ICG with Kolliphor HS 15, the dye was

lyophilized by the manufacturer (mivenion GmbH). Lyophilized

ICG/HS 15 was stored at 28 to 68C. Both dyes were

reconstituted on the day of injection to a concentration of 1

mg/mL with distilled water (Ampuwa; Fresenius Kabi, Bad

Homburg, Germany).

A low, medium, and high dose (0.05, 0.1, and 0.15 mg/kg

body weight [b.w.], respectively) for conventional ICG and

ICG/HS 15 were each tested in five individual animals (n ¼ 30),

respectively (Tables 1, 2). Each animal underwent serial

intravenous injection of the identical dye and concentration

followed by in vivo fluorescence imaging at days 7, 14, and 21.

For more details, please use the following link:

https://www.google.ps/url?sa=t&rct=j&q=&esrc=s&source=web&cd=2&cad=rja&uact=8&ved=0ahUKEwiW5cmcutzKAhUGvBoKHetEDGoQIAgiMAE&url=http%3A%2F%2Fwebcache.googleusercontent.com%2Fsearch%3Fq%3Dcache%3A9TNK6utCHe0J%3Ajov.arvojournals.org%2Fdata%2FJournals%2FIOVS%2F933256%2Fi1552-5783-55-10-6204.pdf%2B%26cd%3D2%26hl%3Den%26ct%3Dclnk%26gl%3Dps&usg=AFQjCNGKKcjMvVxZj3M7rlsyb_8U8DCWUg&sig2=tqAl-T5lvA2sZJ5klaPrbg

file:///C:/Users/Rafik%20Karaman/Downloads/i1552-5783-55-10-6204.pdf

Hoping this will be helpful,

Rafik