Devices and method for enrichment and alteration of cells and other particles
The invention features devices and methods for the deterministic separation of particles. Exemplary methods include the enrichment of a sample in a desired particle or the alteration of a desired particle in the device. The devices and methods are advantageously employed to enrich for rare cells, e.g., fetal cells, present in a sample, e.g., maternal blood and rare cell components, e.g., fetal cell nuclei. The invention further provides a method for preferentially lysing cells of interest in a sample, e.g., to extract clinical information from a cellular component, e.g., a nucleus, of the cells of interest. In general, the method employs differential lysis between the cells of interest and other cells (e.g., other nucleated cells) in the sample.
1. A method of enriching a sample in fetal cells relative to maternal cells, the method comprising:
(a) introducing a maternal blood sample into a microfluidic device capable of enriching fetal nucleated cells relative to maternal cells based on size, shape, deformability, or affinity to produce an enriched sample;
(b) lysing all cells in the enriched sample to release nuclei;
(c) performing telomere length analysis to identify fetal nuclei;
(d) collecting fetal nuclei; and
(e) analyzing fetal nuclei.
2. The method of claim 1 , wherein step (d) comprises collecting fetal nuclei by selectively lysing nuclei of unwanted cells relative to fetal nuclei.
3. The method of claim 1 , wherein the microfluidic device comprises a channel having a structure that deterministically directs fetal nucleated cells in a first direction and at least some maternal cells in a second direction based on deterministic lateral displacement.
4. The method of claim 3 , wherein the microfluidic device is a duplex device comprising a channel comprising a first section comprising first and second outer regions, each outer region comprising a structure that deterministically directs particles having a hydrodynamic size above a critical size in a first direction and particles having a hydrodynamic size below the critical size in a second direction, wherein the first and second outer regions are aligned in parallel in the channel.
5. The method of claim 1 , wherein step (e) comprises employing RNA Fluorescence In Situ Hybridization (FISH) for positive or negative selection of fetal nuclei.
6. The method of claim 1 , wherein step (e) comprises employing sequencing for positive or negative selection of fetal nuclei.
7. The method of claim 1 , wherein step (e) comprises analyzing fetal nuclei by micro-dissection.
8. The method of claim 1 , wherein step (e) comprises analyzing fetal nuclei by polymerase chain reaction (PCR) or whole genome amplification (WGA).
9. The method of claim 1 , wherein step (e) comprises analyzing fetal nuclei by analysis of short tandem repeats (STR).
10. The method of claim 1 , wherein step (e) comprises analyzing fetal nuclei by analysis of single nucleotide point mutations (SNP), deletions, or translocations.
11. The method of claim 1 , wherein step (e) comprises analyzing fetal nuclei by detecting chromosomal aneuploidies.
12. The method of claim 11 , wherein chromosomal aneuploidies are detected using comparative genome hybridization.
13. A method of enriching a sample in fetal cells relative to maternal cells, the method comprising:
(a) introducing a maternal blood sample into a microfluidic device capable of enriching fetal nucleated cells relative to maternal cells based on size, shape, deformability, or affinity to produce an enriched sample;
(b) lysing all cells in the enriched sample to release nuclei;
(c) selectively lysing nuclei of unwanted cells and extracting fetal apoptotic DNA from fetal nuclei; and
(d) analyzing the extracted fetal apoptotic DNA.
14. The method of claim 13 , wherein the microfluidic device comprises a channel having a structure that deterministically directs fetal nucleated cells in a first direction and at least some maternal cells in a second direction based on deterministic lateral displacement.
15. The method of claim 14 , wherein the microfluidic device is a duplex device comprising a channel comprising a first section comprising first and second outer regions, each outer region comprising a structure that deterministically directs particles having a hydrodynamic size above a critical size in a first direction and particles having a hydrodynamic size below the critical size in a second direction, wherein the first and second outer regions are aligned in parallel in the channel.
16. The method of claim 13 , wherein step (d) comprises analyzing the extracted fetal apoptotic DNA by polymerase chain reaction (PCR) or whole genome amplification (WGA).
17. The method of claim 13 , wherein step (d) comprises analyzing the extracted fetal apoptotic DNA by analysis of short tandem repeats (STR).
18. The method of claim 13 , wherein step (d) comprises analyzing the extracted fetal apoptotic DNA by analysis of single nucleotide point mutations (SNP), deletions, or translocations.
19. The method of claim 13 , wherein step (d) comprises analyzing the extracted fetal apoptotic DNA by detecting chromosomal aneuploidies.
20. The method of claim 19 , wherein chromosomal aneuploidies are detected using comparative genome hybridization.