Granted Patent B2

US 12,566,113 · App. 17/845,331 · Granted Mar 3, 2026

Inventors: Joshua Delaney (Pleasanton, CA); Shalini Gohil (Pleasanton, CA); Adam Lowe (Pleasanton, CA); Yi Luo (Pleasanton, CA); Dagmar Walter (Pleasanton, CA)

Assignee: 10x Genomics, Inc.

G01N1/30C07D403/12G01N2001/305

Abstract

Claims (28)

1 . A method for preparing a biological sample comprising contacting the sample with a first fixing reagent composition comprising:

a compound of formula (I)

wherein, “Linker” is a linear alkane moiety of 2-24 carbons; a linear alkane moiety of 2-24 carbons comprising a disulfide bond; or a linear alkane moiety of 2-24 carbons substituted with —OR, wherein R is selected from —H, —O(CO)—CH 3 , and —O(CO)— imidazole; and m is 1 to 12; and

a carrier.

2 . The method of claim 1 , wherein the compound of formula (I) is a compound of formula (II)

wherein n is 1 to 13.

3 . The method of claim 2 , wherein the compound of formula (II) is selected from compounds (2a)-(2k)

4 . The method of claim 1 , wherein the compound of formula (I) is a compound of formula (IV)

wherein m is 1 to 11, and n is 1 to 11.

5 . The method of claim 4 , wherein the compound of formula (IV) is selected from compounds (4a)-(4d)

6 . The method of claim 1 , wherein the compound of formula (I) is compound of formula (V)

wherein R is selected from —H, —O(CO)—CH 3 , and —O(CO)— imidazole.

7 . The method of claim 6 , wherein the compound of formula (V) is selected from compounds (5a)-(5c)

8 . The method of claim 1 , wherein the first fixing reagent composition comprises:

(a) the compound of formula (I) at a concentration of 5 mM to 50 mM; and

(b) DMSO at a concentration of 2.5% to 5%.

9 . The method of claim 1 , wherein the method further comprises contacting the sample with a second fixing reagent, wherein the second fixing reagent comprises:

i) paraformaldehyde;

ii) a compound of formula (III)

wherein n is 1 to 12; or

iii) a compound of formula (V)

wherein R is selected from —H, —O(CO)—CH 3 , and —O(CO)— imidazole.

10 . The method of claim 1 , wherein the method further comprises contacting the sample with a lysis agent and/or an unfixing agent, wherein the lysis agent comprises a compound capable of cleaving a carbamate bond or a compound capable of cleaving a disulfide bond and the unfixing agent comprises a compound capable of reversing paraformaldehyde fixation, wherein:

the compound capable of cleaving a carbamate bond is DETA, EDA, hydrazine monohydrate, a carboxyesterase, or a combination thereof;

the compound capable of cleaving a disulfide bond is DTT; and

the compound capable of reversing paraformaldehyde fixation is selected from any one of compounds (7a)-(7o)

11 . The method of claim 1 , wherein the biological sample is a tissue sample.

12 . The method of claim 5 , wherein the compound of formula (IV) is compound (4a).

Assignments (1)

Continuity (4)

References Cited (400)

Cited By (3)

US 12,637,706 US 12,668,835 US 12,692,492