Methods of chromosome drying and spreading
The present invention provides for a method of drying and spreading chromosomes from various biological samples to yield optimal chromosomal spreading. The method requires preparing a biological sample for treatment, providing a cytogenetic chamber capable of setting predetermined conditions, pre-testing a portion of the biological sample in the cytogenetic chamber, and finally treating the remaining biological sample. The method is useful to yield metaphase chromosomes that are small and rounded, with very few overlapping or scattered chromosomes. Furthermore, the method is uses restricted ranges of temperature and relative humidity to achieve consistent chromosomal spreading. The morphologies of the chromosomes are preserved in order to execute banding techniques at 550 bands and chromosomal analysis on high-resolution chromosomes.
1. A method of drying chromosomes, the method comprising the steps of:
a) preparing a biological sample for treatment by:
i) obtaining a biological sample of cells for treatment;
ii) transferring the sample of cells onto a support;
iii) harvesting the sample of cells;
iv) adding a fixative agent to the cells after the cells are harvested;
b) providing a cytogenetic chamber capable of setting predetermined conditions;
c) pre-testing the cytogenetic chamber by:
i) setting the chamber to a predetermined condition based on the biological sample being treated;
ii) placing a pre-testing portion of the cells with the fixative agent into the chamber and reserving a remainder of the cells;
iii) allowing the pre-testing cells to become acclimated to the predetermined conditions before drying the pre-testing cells;
iv) aspirating all of the fixative from the pre-testing cells;
v) drying the pre-testing cells until spreading is optimized for consistency;
vi) removing the pre-testing cells and analyzing for optimal chromosome spreading for consistency;
d) placing the reserved cells into the chamber at the same predetermined conditions used for the pre-testing, allowing the reserved cells to become acclimated to the predetermined conditions before drying the reserved cells, aspirating the fixative from the reserved cells, drying the reserved cells, and analyzing the reserved cells for optimal chromosomal spreading.
2. The method of claim 1 , wherein the predetermined condition of the chamber has a temperature of about 30.0-32.0° C. and a relative humidity of about 35-37%.
3. The method of claim 2 , wherein the biological sample comprises an amnioctye, an early amniocyte, and a chorionic villus sampling (CVS).
4. The method of claim 1 , wherein the predetermined condition of the chamber has a temperature of about 31.0-33.0° C. and a relative humidity of about 36-38%.
5. The method of claim 4 , wherein the biological sample comprises a product of conception or skin tissue.
6. The method of claim 1 , wherein the predetermined condition of the chamber has a temperature of about 32° C. and a relative humidity of about 36-46%.
7. The method of claim 1 , wherein the biological sample comprises a blood specimen, a bone marrow, and a tumor cell.
8. The method of claim 1 , wherein the step of harvesting isolates the cells to form a solution of viable cells.
9. The method of claim 1 , wherein the fixture agent comprises a 3:1 ratio of methanol to glacial acetic acid.
10. The method of claim 1 , wherein the step of allowing the pre-testing cells to become acclimated to the predetermined conditions take about 15 minutes.
11. The method of claim 1 , wherein the step of aspirating the fixative from the pre-testing cells requires about 3 seconds.
12. The method of claim 1 , wherein the step of drying the pre-testing cells requires about 2 minutes.
13. A method of drying chromosomes, the method comprising the steps of:
a) preparing a biological sample for treatment by:
i) obtaining a biological sample of cells for treatment, wherein the biological sample comprises an amnioctye, an early amniocyte, and a chorionic villus sampling (CVS);
ii) transferring the sample of cells onto a support;
iii) harvesting the sample of cells;
iv) adding a fixative agent to the cells after the cells are harvested;
b) providing a cytogenetic chamber capable of setting predetermined conditions;
c) pre-testing the cytogenetic chamber by:
i) setting the chamber to a predetermined condition, wherein the predetermined condition is a temperature of about 30.0-32.0° C. and a relative humidity of about 35-37%;
ii) placing a pre-testing portion of the cells with the fixative agent into the chamber and reserving a remainder of the cells;
iii) allowing the pre-testing cells to become acclimated to the predetermined conditions before drying the pre-testing cells;
iv) aspirating all of the fixative from the pre-testing cells;
v) drying the pre-testing cells until spreading is optimized for consistency;
vi) removing the pre-testing cells and analyzing for optimal chromosome spreading for consistency;
d) placing the reserved cells into the chamber at the same predetermined conditions used for the pre-testing, allowing the reserved cells to become acclimated to the predetermined conditions before drying the reserved cells, aspirating the fixative from the reserved cells, drying the reserved cells, and analyzing the reserved cells for optimal chromosomal spreading.
14. A method of drying chromosomes, the method comprising the steps of:
a) preparing a biological sample for treatment by:
i) obtaining a biological sample of cells for treatment, wherein the biological sample comprises a product of conception and skin tissue;
ii) transferring the sample of cells onto a support;
iii) harvesting the sample of cells;
iv) adding a fixative agent to the cells after the cells are harvested;
b) providing a cytogenetic chamber capable of setting predetermined conditions;
c) pre-testing the cytogenetic chamber by:
i) setting the chamber to a predetermined condition, wherein the predetermined condition is a temperature of about 31.0-33.0° C. and a relative humidity of about 36-38%;
ii) placing a pre-testing portion of the cells with the fixative agent into the chamber and reserving a remainder of the cells;
iii) allowing the pre-testing cells to become acclimated to the predetermined conditions before drying the pre-testing cells;
iv) aspirating all of the fixative from the pre-testing cells;
v) drying the pre-testing cells until spreading is optimized for consistency;
vi) removing the pre-testing cells and analyzing for optimal chromosome spreading for consistency;
d) placing the reserved cells into the chamber at the same predetermined conditions used for the pre-testing, allowing the reserved cells to become acclimated to the predetermined conditions before drying the reserved cells, aspirating the fixative from the reserved cells, drying the reserved cells, and analyzing the reserved cells for optimal chromosomal spreading.
15. A method of drying chromosomes, the method comprising the steps of:
a) preparing a biological sample for treatment by:
i) obtaining a biological sample of cells for treatment, wherein the biological sample comprises a blood specimen, a bone marrow, and a tumor cell;
ii) transferring the sample of cells onto a support;
iii) harvesting the sample of cells;
iv) adding a fixative agent to the cells after the cells are harvested;
b) providing a cytogenetic chamber capable of setting predetermined conditions;
c) pre-testing the cytogenetic chamber by:
i) setting the chamber to a predetermined condition, wherein the predetermined condition is a temperature of about 32° C. and a relative humidity of about 36-46%;
ii) placing a pre-testing portion of the cells with the fixative agent into the chamber and reserving a remainder of the cells;
iii) allowing the pre-testing cells to become acclimated to the predetermined conditions before drying the pre-testing cells;
iv) aspirating all of the fixative from the pre-testing cells;
v) drying the pre-testing cells until spreading is optimized for consistency;
vi) removing the pre-testing cells and analyzing for optimal chromosome spreading for consistency;
d) placing the reserved cells into the chamber at the same predetermined conditions used for the pre-testing, allowing the reserved cells to become acclimated to the predetermined conditions before drying the reserved cells, aspirating the fixative from the reserved cells, drying the reserved cells, and analyzing the reserved cells for optimal chromosomal spreading.