Integrated analysis device analysis techniques
Provided are integrated analysis devices having features of macroscale and nanoscale dimensions, and devices that have reduced background signals and that reduce quenching of fluorophores disposed within the devices. Related methods of manufacturing these devices and of using these devices are also provided.
1. A method of analysis, comprising:
translocating a DNA molecule through a primary fluidic channel and two or more additional fluidic channels in fluid communication with the primary fluidic channel and of successively decreasing width such that at least a portion of the DNA molecule is elongated while disposed in a nanochannel analysis region of the two or more additional fluidic channels, wherein the total widths of the narrowest additional fluidic channels are approximately equal to the width of the primary fluidic channel;
detecting a signal arising from the elongated DNA molecule or portion thereof within the nanochannel analysis region; and
correlating the signal to a property of the DNA molecule.
2. The method of claim 1 , wherein the primary fluidic channel has a width in the range of from about 10 nm to less than about 10000 nm.
3. The method of claim 1 , wherein the primary fluidic channel has a width in the range of from about 100 nm to less than about 1000 nm.
4. The method of claim 1 , wherein the primary fluidic channel has a depth in the range of from about 10 nm to less than about 10000 nm.
5. The method of claim 1 , wherein the primary fluidic channel has a depth in the range of from about 100 nm to less than about 1000 nm.
6. The method of claim 1 , wherein the ratio of the widths of the primary fluidic channel and each of the narrowest additional fluidic channels is in the range of from about 100 to about 10000.
7. The method of claim 1 , wherein the translocating is accomplished by application of an electrical gradient, a pressure gradient, a magnetic field, a thermal gradient, or any combination thereof.
8. The method of claim 1 , further comprising translocating the DNA molecule through at least two channels of successively increasing width.
9. The method of claim 1 , further comprising reversing the direction of a gradient so as to reverse the direction of the DNA molecule such that at least a portion of the DNA molecule re-enters a channel.
10. The method of claim 9 , further comprising detecting a signal from the DNA molecule after re-entering the channel.
11. The method of claim 1 , wherein the nanochannel analysis region comprises nanochannels of varying depth, varying width, or varying depth and width.
12. The method of claim 1 , wherein the nanochannel analysis region comprises a nanochannel having a width of 1 nm to 1000 nm.
13. The method of claim 1 , wherein the nanochannel analysis region comprises a nanochannel having a width of 10 nm to 500 nm.
14. The method of claim 1 , wherein the nanochannel analysis region comprises a nanochannel having a depth of 10 nm to 100 nm.
15. The method of claim 1 , wherein the nanochannel analysis region comprises a nanochannel having at least one linear segment having a length in the range of from about 0.1 microns to about 50 microns.
16. The method of claim 1 , wherein the nanochannel analysis region comprises a nanochannel comprising at least one bend of at least about 30 degrees.
17. The method of claim 1 , wherein the detecting is accomplished optically, electrically, magnetically, electromagnetically, or any combination thereof.
18. The method of claim 1 , wherein the DNA molecule comprises a label.
19. The method of claim 18 , wherein the label comprises a fluorophore, a quantum dot, a metal dot, a polymer bead, a lanthanide chelate, a nanoparticle, a fluorescent bead, a phosphorescent bead, a semiconductor nanoparticle, a dendrimer, a molecular antennae, or any combination thereof.
20. The method of claim 1 , comprising illuminating the DNA molecule with electromagnetic radiation having a wavelength in the range of from about 10 nm to about 2500 nm.