Good morning,

I want to dissolve cholesterol in Krebs-Henseleit buffer in tissue organ bath chamber where I place tracheal tissue. Krebs-H. buffer is buffer composed of water and salts similar to Ringer buffer. I want to add cholesterol to the chamber as a potential masking agent of the tracheal receptors. I followed this protocol to make cholesterol soluble in water, however the cholesterol did not dissolve after the protocol was done and cholesterol looks undissolved (photo attached). This is the protocol I followed:

Cholesterol was dissolved in a 1:1:0.9 ratio by volume of chloroform, methanol, and distilled water, respectively. The fluids were vortexed and the top phase of water and methanol was discarded. The bottom phase of chloroform and cholesterol was then transferred into a glass tube and dried under N2. The cholesterol was then resuspended in 0.9% saline. The concentration of the solution was 2.7 mg/ml.

Can anyone suggest how to dissolve cholesterol so I can use it in water solutions?

Thank you.

Juliana