Selective near-infrared optical imaging of necrotic cells and simultaneous cell fixing and counter staining with metallacrown complexes

Patent metadata

Field	Value
Publication number	US-10458991-B2
Application number	US-201615752870-A
Country	US
Kind code	B2
Filing date	Aug 12, 2016
Priority date	Aug 14, 2015
Publication date	Oct 29, 2019
Grant date	Oct 29, 2019

Abstract

In an example of a method for simultaneously fixing and staining cells, the cells are initially incubated in a solution including a Ln(III)Zn16(HA ligand)16 metallacrown complex, wherein the HA ligand is a hydroximate ligand. The incubating cells are exposed to ultraviolet (UV) light. The cells are allowed to continue to incubate in the solution after UV light exposure.

First claim

The invention claimed is:

1. A method for simultaneously fixing and staining cells, the method comprising:
initially incubating the cells in a solution including a Ln(III)Zn 16 (HA ligand) 16 metallacrown complex, wherein the HA ligand is a hydroximate ligand;
exposing the incubating cells to ultraviolet (UV) light; and
continuing to incubate the cells in the solution after UV light exposure.

2. The method of claim 1 wherein:
the solution includes the Ln(III)Zn 16 (HA ligand) 16 metallacrown complex in a medium; and
a concentration of the Ln(III)Zn 16 (HA ligand) 16 metallacrown complex in the solution ranges from about 90 μM to about 400 μM.

3. The method of claim 1 , further comprising culturing the cells prior to incubating the cells in the solution.

4. The method of claim 1 , wherein:
initially incubating the cells is accomplished for a time (T1) ranging from about 10 minutes to about 3 hours;
exposing the incubating cells to UV light is accomplished for a time (T2) ranging from about 5 minutes to about 10 minutes; and
continuing to incubate the cells is accomplished for a time (T3) ranging from about 1 hour to about 2 hours.

5. The method as defined in claim 4 , further comprising exposing the incubating cells to additional UV light for a time (T4) ranging from about 1 minute to about 5 minutes.

6. The method of claim 1 , wherein the HA ligand of the Ln(III)Zn 16 (HA ligand) 16 metallacrown complex is selected from the group consisting of pyrazinehydroximate, quinoxalinehydroximate, quinaldinehydroximate, and combinations thereof.

7. The method as defined in claim 6 wherein:
the Ln(III)Zn 16 (HA ligand) 16 metallacrown complex is one of:
[Ln(III)Zn 16 (pyrazinehydroximate) 16 (pyridine) 8 ] counter ion;
[Ln(III)Zn 16 (quinoxalinehydroximate) 16 (pyridine) 8 ] counter ion; or
[Ln(III)Zn 16 (quinaldinehydroximate) 16 (pyridine) 8 ] counter ion;
wherein the Ln(M) is selected from the group consisting of Y 3+ , La 3+ , Ce 3+ , Pr 3+ , Nd 3+ , Pm 3+ , Sm 3+ , Eu 3+ , Gd 3+ , Tb 3+ , Dy 3+ , Ho 3+ , Er 3+ , Tm 3+ , Yb 3+ , and Lu 3+ ; and wherein the counter ion is selected from the group consisting of a triflate, a mesylate, a besylate, a camsylate, an edisylate, an estolate, an esylate, a napsylate, a tosylate, a fluoride, a chloride, a bromide, an iodide, a nitrate, a sulfate, a carbonate, an acetate, a phosphate, and a sulfonate.

8. The method as defined in claim 6 wherein:
the Ln(III)Zn 16 (HA ligand) 16 metallacrown complex includes a mixture of metallacrown complexes;
each species in the metallacrown mixture is [Ln(III)Zn 16 (pyrazinehydroximate) x (quinoxalinehydroximate) y (pyridine) 8 ] counter ion,
wherein x+y=16;
wherein the Ln(M) is selected from the group consisting of Y 3+ , La 3+ , Ce 3+ , Pr 3+ , Nd 3+ , Pm 3+ , Sm 3+ , Eu 3+ , Gd 3+ , Tb 3+ , Dy 3+ , Ho 3+ , Er 3+ , Tm 3+ , Yb 3+ , and Lu 3+ ; and
wherein the counter ion is selected from the group consisting of a triflate, a mesylate, a besylate, a camsylate, an edisylate, an estolate, an esylate, a napsylate, a tosylate, a fluoride, a chloride, a bromide, an iodide, a nitrate, a sulfate, a carbonate, an acetate, a phosphate, and a sulfonate.

9. The method of claim 8 , wherein x ranges from 8 to 13 and y ranges from 3 to 8.

10. The method of claim 1 , wherein the medium is a serum-supplemented medium.

11. The method of claim 1 , wherein the UV light is UV-A light.

12. The method of claim 1 , wherein:
UV light exposure induces death of at least some of the cells; and
after continuing to incubate the cells, the Ln(III)Zn 16 (HA ligand) 16 metallacrown complex is located in nuclei and cytoplasm of least some dead cells.

Assignees

• Univ Michigan Regents
• Centre Nat Rech Scient

Inventors

• Martinic Ivana
• Nguyen Tu Ngoc
• Eliseeva Svetlana V
• Pecoraro Vincent L
• Petoud Stéphane

Classifications

• G01N33/583 Primary

  with non-fluorescent dye label · CPC title

• C07F3/06

  Zinc compounds · CPC title

• G01N21/6458

  Fluorescence microscopy (fluorescence microscopes per se G02B21/0076 and G02B21/16) · CPC title

• G01N2001/302

  Stain compositions · CPC title

• G01N2001/305

  Fixative compositions · CPC title