Table 2. Properties of the optical clearing agents and clearing procedures utilized within the present study.

					Processing + Clearing Time3
Clearing (and Mounting) Agents	Concentration1 and Molecular Weight (g/mol) of Final Clearing Agent	RI2	Aqueous vs. Non-Aqueous	Single- or Multi-step Procedure	Murine Bone Segments	Intact Murine Knee Joints	Cost of Clearing Agents4 ($US/mL)	References
Water5	100% Water (MW 18.02)	1.34	Aq	Single	Not Applicable	Not Applicable	Not Applicable
Visikol	100% Visikol (Makeup and MWs are Proprietary)	1.45	Aq	Single	~24-hrs	4-d	$1.20	www.visikol.com
Clear T2	50% formamide (MW 45.04) + 20% PEG (MW ~8000)	1.45	Aq	Multi	~48-hrs	6-d	$0.41	[39]
Focus Clear	100% Focus Clear (Makeup and MWs are Proprietary)	1.46	Aq	Single	2-d	6-d	$78.00	www.cedarlanelabs.com
2, 2'-thiodiethanol (TDE)	97% TDE (MW 122.19) in water	1.47	Aq	Multi	2-d	4-d	$1.04	[41]
See Deep Brain (SeeDB)	80.2% D-fructose wt/wt (MW 180.16) in water	1.50	Aq	Multi	4-d	14-d	$0.29	[40]
Methyl salicylate (MS)	100% MS (MW 152.15)	1.51	Non-Aq	Multi	4-d	7-d	$0.31	[58]
Benzyl benzoate-benzyl alcohol (BABB)	25% benzyl alcohol (MW 108.14) + 75% benzyl benzoate (MW 212.24)	1.53	Non-Aq	Multi	4-d	7-d	$0.30	[43]
Tetrahydrofuran-dibenzyl ether (THF-DBE)	100% DBE (MW 198.26)	1.56	Non-Aq	Multi	4-d	7-d	$0.16	[42]
Acrylic6	85% methyl methacrylate (MW 100.12) + 15% n-butyl phthalate (MW 278.34) + 2% benzoyl peroxide (wt/vol) (MW 242.23)	1.49	Non-Aq	Multi	9-d	14-d	$1.13	[26]

¹All concentrations are indicated as vol/vol unless otherwise noted

²RI values obtained from literature

³Values indicate approximate total time required for full processing and clearing protocol

⁴Reference provided is for the cost of clearing reagents only at the time of the study; cost of common processing reagents (e.g. ethanol, etc.) was not included

⁵Water refers to non-cleared samples immersed in deionized water

⁶Acrylic refers to the plastic embedding procedure; samples are infiltrated with the indicated agents and then polymerized within the solid support matrix