Fixative solutions and methods for using same

An in vitro method for fixating at least one cell, wherein said method comprises the step of

• a) contacting the at least one cell with a liquid, a compound of Formula I, and a compound of Formula II, so as to form a mixture;
• wherein the concentration of the compound of Formula I in said mixture is in a range of from 0.8 mM to 2.4 mM;
• wherein the concentration of the compound of Formula II in said mixture is sufficient to dissolve substantially all of the compound of Formula I in said mixture;
• wherein the compound of Formula I is:
• wherein the compound of Formula II is:
• wherein R1, R2, and R3 are each independently selected from the group consisting of —C(O)O—N-succinimide, —C(O)F, —C(O)Cl, —C(O)Br, —C(O)—N3, —C(O)O-pentafluorophenyl, —C(O)O—N-benzotriazole, —C(O)O-phthalimide, —C(O)—O—C(O)CH3, —NCS, —NCO, —S(O)2Cl, —S(O)2F, phenylsulfonyl fluoride, phenylsulfonyl chloride, and C2 epoxidyl;
• wherein X1, X2, and X3 are each independently selected from the group consisting of C1-C6 alkylene, and phenylene;
• wherein Q is a bond or C1-6 alkylene;
• wherein Z is heteroaryl or aryl;
• and
• wherein each alkylene, phenylene, heteroaryl, and aryl is optionally substituted.

A method for preparing a fixative solution comprising a compound of Formula I and a compound of Formula II, wherein said method comprises the steps of:

• a) contacting a compound of Formula I with a solvent, so as to form a first solution;
• b) contacting said first solution with a compound of Formula II, so as to form second solution;
• c) adding a buffer solution, to the second solution, so as to form the fixative solution;
• wherein the concentration of the compound of Formula II in said fixative solution is sufficient to dissolve substantially all of the compound of Formula II in said fixative solution;
• wherein the second solution is subjected to mixing, during or upon the addition of the buffer solution;
• wherein the buffer solution has a temperature in a range of from 15° C. to 45° C., preferably in a range of from 35° C. to 40° C.;
• wherein the compound of Formula I is:
• wherein the compound of Formula II is:
• wherein R1, R2, and R3 are each independently selected from the group consisting of —C(O)O—N-succinimide, —C(O)F, —C(O)Cl, —C(O)Br, —C(O)—N3, —C(O)O-pentafluorophenyl, —C(O)O—N-benzotriazole, —C(O)O-phthalimide, —C(O)—O—C(O)CH3, —NCS, —NCO, —S(O)2Cl, —S(O)2F, phenylsulfonyl fluoride, phenylsulfonyl chloride, and C2 epoxidyl;
• wherein X1, X2, and X3 are each independently selected from the group consisting of C1-C6 alkylene, and phenylene;
• wherein Q is a bond or C1-6 alkylene;
• wherein Z is heteroaryl or aryl;
• and
• wherein each alkylene, phenylene, heteroaryl, and aryl is optionally substituted.